Methods and compositions of novel triazine compounds

ABSTRACT

The present invention relates to methods and compositions comprising compounds that treat pathophysiological conditions arising from inflammatory responses. In particular, the present invention is directed to compounds that inhibit or block glycated protein produced induction of the signaling-associated inflammatory response in endothelial cells. The present invention relates to compounds that inhibit smooth muscle proliferation. In particular, the present invention is directed to compounds that inhibit smooth muscle cell proliferation by modulating HSPGs such as Perlecan. The present invention further relates to the use of compounds to treat vascular occlusive conditions characterized by smooth muscle proliferation such as restenosis and atherosclerosis.

PRIOR RELATED U.S. APPLICATION DATA

[0001] This application is a continuation-in-part of U.S. patentapplication Ser. No. ______ filed Mar. 17, 2003, which is a continuationof U.S. patent application Ser. No. 10/253,388 filed Sep. 23, 2002,which claims priority to U.S. Provisional Application Serial Number60/324,147 filed Sep. 21, 2001, which is incorporated herein byreference.

FIELD OF THE INVENTION

[0002] The present invention relates to triazine compounds. Moreparticularly, the invention relates to methods and compositions formaking and using triazine compounds.

BACKGROUND OF THE INVENTION

[0003] Synthesis of novel compounds leads to new possibilities fordiscovery of novel therapeutic interventions. By using structure andactivity relationship investigations, compounds can be tailored so thatthe compounds have at least one activity that can be predicted from itsstructure. Using high-throughput assays allows for the rapiddetermination of the activity of the newly synthesized compounds.

[0004] Novel compounds for new therapeutic interventions are needed formany areas of medicine and disease treatment. For example, chronic andacute inflammatory conditions form the basis for diseases affecting allorgan systems including, but not limited to, asthma, acute inflammatorydiseases, vascular inflammatory disease, chronic inflammation,atherosclerosis, angiopathy, myocarditis, nephritis, Crohn's disease,arthritis, type I and II diabetes and associated vascular pathologies.The incidence of these inflammatory conditions is on the rise in thepopulation as a whole, with diabetes alone affecting 16 million people.

[0005] While inflammation in and of itself is a normal immune response,chronic inflammation leads to complications and ongoing system damagedue to the interactions of unknown cellular factors. In particular,chronic inflammation can cause endothelial damage resulting in vascularcomplications. Coronary artery, cerbrovascular and peripheral vasculardisease resulting from atherosclerotic and thromboembolicmacroangiopathy are the primary causes of mortality in chronicinflammatory diseases.

[0006] Many humans and animals have limited lifespans and lifestylesbecause of conditions relating to lifestyle choices, such as diet andexercise, or because of genetic predispositions to develop a disease.For example, vascular smooth muscle cell proliferation is a commonconsequence of endothelial injury and is believed to be an earlypathogenetic event in the formation of atherosclerotic plaques orcomplications related to vascular injury or as a result surgicalinterventions. Abnormal vascular smooth muscle cell (SMC) proliferationis thought to contribute to the pathogenesis of vascular occlusivelesions, including arteriosclerosis, atherosclerosis, restenosis, andgraft atherosclerosis after organ transplantation.

[0007] Percutaneous coronary artery intervention (PTCA) procedures arethe most common in-patient hospital procedure in the United States.According to the American Heart Association, about one-third of thepatients that undergo balloon angioplasty have restenosis of the widenedsegment of the vessel within approximately 6 months. It may be necessaryto perform another angioplasty or coronary artery bypass surgery onrestenosed arteries. A key feature of restenosis is an injury responsethat results in activation of an inflammatory cascade and remodeling ofthe cells both inside and outside the carotid artery wall. This includesexcessive growth of connective tissue and smooth muscle into the lumenof the artery known as neointimal hyperplasia. Currently there are noeffective pharmacological treatments available that control thepathogenesis of vascular occlusive lesions, such as, but not limited to,arteriosclerosis, atherosclerosis, restenosis, and graft atherosclerosisafter organ transplantation. Identification of effective therapeuticswith minimal side effects will restore quality of life without requiringadditional surgical procedures such as coronary artery bypass surgery.

[0008] Control or modulation of factors produced by the body in responseto injury, surgery, metabolic factors or loss of control of in feedbackmechanisms, leading to too much or too little of a factor has long beenthe goal of administering pharmacological agents. One disease thatrapidly growing in the industrialized countries is the occurrence ofdiabetes and all of its attendant sequellae. One of the factorsimportant in the damage associated with diabetes is the presence ofglycated proteins.

[0009] Glycated proteins and advanced glycation end products (AGE)contribute to cellular damage, particularly, diabetic tissue injury, byat least by two major mechanisms; modulation of cellular functionsthrough interactions with specific cell surface receptors, andalteration of the extracellular matrix leading to the formation ofprotein cross-links. Studies suggest that glycated protein and AGEinteractions with cells may promote inflammatory processes and oxidativecellular injury. AGE increases lipoprotein oxidisability andatherogenicity. Its binding to matrix proteins induces synthesis ofcytokines and activates cellular messangers. Diseases where glyeatedprotein and AGE accumulation is a suspected etiological factor includevascular complications of diabetes, microangiopathies, renalinsufficiency and Alzheimer's disease.

[0010] The exact mechanisms by which high plasma glucose, as seen indiabetes, causes microvascular damage are not completely understood. Onepotential mechanism by which hyperglycemia can be linked tomicroangiopathies is through the process of non-enzymatic glycation ofcritical proteins. Non-enzymatic glycation, i.e. the linking of proteinswith glucose, leads to the formation of glycated proteins. The firststep in this glycation pathway involves the non-enzymatic condensationof glucose with free amino groups in the protein, primarily theepsilon-amino groups of lysine residues, forming the Amadori adducts.These early glycation products can undergo further reactions such asrearrangements, dehydration and condensations to form irreversibleadvanced glycation end products (AGE). These are a highly reactive groupof molecules whose interaction with specific receptors on thecell-surface which are thought to lead to pathogenic outcomes.

[0011] Other major area of disease of where treatments are needed andfor which adequate and effective therapies do not exist are cellularproliferative disorders, or disorders caused by unwanted or unintendedcellular growth. As mentioned, smooth muscle cell (SMC) hyperplasia is amajor event in the development of atherosclerosis and is alsoresponsible for the significant number of failure rates followingvascular procedures such as angioplasty, stent implantation and coronaryartery bypass surgery. In the normal vessel, SMC are quiescent, but theyproliferate when damage to the endothelium occurs. Naturally occurringgrowth modulators, many of which are derived from the endothelium,tightly control SMC proliferation in vivo. When the control becomesunregulated, a pathological state is induced in the subject.

[0012] Another major area of unwanted cellular growth, that is uncheckedby the body's regulatory systems, is cancer or oncological conditions.Many therapies have been used and are being used in an effort to restorehealth or at least stop the unwanted cell growth. Many times,therapeutic agents can have an effect individually, but often,therapeutic regimes require combinations of different pharmacologicalagents with treatments such as surgery or radiation.

[0013] There is a present need for treatments of chronic or acutediseases, such as atherosclerosis, unwanted cellular growth or cellularproliferation, diabetes, inflammatory conditions and vascular occlusivepathologic conditions, because occurrence is frequent, the currentlyavailable treatments are costly and the conditions are refractory tomany pharmacological therapies. The mechanisms involved in the controlor prevention of such diseases are not clear and there exists a need forpreventive and therapeutic treatments of these and other diseases Thus,what is presently needed are novel compounds that find utility inmethods and compositions for treatment and prevention of chronic andacute diseases.

SUMMARY OF THE INVENTION

[0014] The present invention is directed to methods and compositionscomprising novel compounds, primarily based on a substituted triazinecore. Disclosed herein are methods for making novel compounds, thecompounds, compositions comprising the compounds, and methods andcompositions for using the compounds. The compounds and compositionscomprising the compounds have utility in treatment of a variety ofdiseases.

[0015] Compositions in accordance with the present invention comprisetriazine compounds, analogs, derivatives, and mixtures thereof. Suchtriazine compounds comprise the following structure, where N^(A), N^(B)and N^(C) are typically used to represent pendant substituted aminogroups attached to 1,3,5-triazine at the 2, 4 and 6 positions:

[0016] An example of such triazine compounds includes compounds havingthe following structure.

[0017] In this example, each pendent amino (NRR′) group can representsimply an NH₂ group or a secondary or tertiary amino group, including acyclic secondary amide, and a range of other substituents as describedherein. Compositions in accordance with the present invention alsocomprise analogs of the tris(amino) compounds, that include intermediatecompounds in the synthesis of the tris(amino) triazine compoundsindicated above, for example diamino chlorotriazine compounds, or aminodiclorotriazine compounds shown below, where N^(A) and N^(B) are pendantsubstituted amino groups as described above.

[0018] Compositions in accordance with the present invention alsocomprise analogs of the tris(amino) triazine compounds indicated above,including compounds that are isolated as byproducts in the synthesis ofthe tris(amino) triazine compounds, such as bis(amino)alkoxy triazinecompounds as shown below, where E=O or S and the like.

[0019] The present invention also comprises compositions used in makingthe novel compounds and methods of making the novel compounds disclosedherein.

[0020] The present invention is directed to methods and compositionscomprising compounds that have utility in treatment of pathologicalconditions. One aspect of the present invention comprises compounds andcompositions comprising such compounds in methods for treating diseasesrelated to unwanted cellular proliferation. Many vascular diseases, suchas cardiovascular diseases, organ transplant sequellae, vascularocclusive conditions including, but not limited to, neointimalhyperplasia, restenosis, transplant vasculopathy, cardiac allograftvasculopathy, atherosclerosis, and arteriosclerosis are caused by orhave collateral damage due to unwanted cellular proliferation, such assmooth muscle cell (SMC) hyperplasia. At least one activity of one ormore of these compounds is that the compound has the activity ofeffecting the synthesis of proteoglycans including induction andsynthesis of proteoglycans and active fragments of proteoglycans.Methods comprise administration of compositions comprising compoundsthat have at least the activity of effecting cellular proliferation andeffecting proteoglycan synthesis and activity.

[0021] The present invention also comprises methods and compositionscomprising compounds described herein that have an activity associatedwith modulation of glycosidase enzymes and thus, effecting thesubstrates for such enzymes. Glycosidase enzymes and their activity withtheir substrates, such as proteoglycans or glycated proteins, areaspects of a variety of diseases such as vascular conditions,proteoglycan-associated diseases, kidney disease, autoimmune disease andinflammatory diseases. Compounds described herein that have an activitythat effects the concentrations of substrates of glycosidase enzymes areused in methods of treatment of such vascular, inflammatory, metastaticand systemic diseases.

[0022] An embodiment of the present invention comprises methods andcompositions comprising compounds of the present invention for thetreatment and prevention of conditions or diseases that have as anaspect of the disease or condition, inflammation. An aspect of thepresent invention is directed to methods and compositions comprisingcompounds that are effective in inhibiting inflammation, particularlyinflammation associated with the accumulation or presence of glycatedproteins or AGE. Methods of treatment comprise administration ofcompositions comprising having compounds having at least the activity ofmodulating inflammatory reactions that are components of biologicalconditions including, but not limited to, vascular complications of typeI and type II diabetic-induced vasculopathies, other vasculopathies,microangiopathies, renal insufficiency, Alzheimer's syndrome, andinflammation-induced diseases such as atherosclerosis. An aspect of thepresent invention comprises methods and compositions for the treatmentof diseases, preconditions or pathologies associated with inflammatorycytokines and other inflammation related molecules.

[0023] Another embodiment of the present invention comprises methods andcompositions comprising compounds that have at least the activity ofcausing cellular death or a cessation of cellular activity, referred toherein as cytotoxic activity. This activity can be used in methods forin vitro or in vivo cytotoxicity. For example, compounds having thisactivity can be selectively delivered to an area within a livingorganism to selectively kill cells in that area. Such methods are usingin treating hyperproliferative cells, such as cancers, or other unwantedcellular growth or cellular activities. One aspect of the inventionprovides compositions comprising compounds that nonselectively killcells. Another aspect of the invention provides compounds thatselectively kill cells, for example, cells that have a particularcellular marker or other identifying characteristic such as metabolicrate or uptake of a particular compound.

[0024] The present invention also comprises pharmaceutical compositionscomprising the compounds disclosed herein. Routes of administration anddosages of effective amounts of the compounds and pharmaceuticalcompositions are also disclosed. For example, the compounds of thepresent invention can be administered in combination with otherpharmaceutical agents in a variety of protocols for effective treatmentof disease.

[0025] In another aspect, the present invention relates to drugdelivering or eluting medical devices that contain or are coated with atleast one compound disclosed herein. The medical device suitable for usewith the compounds of the present invention include, but are not limitedto, stents and other medical devices that can provide a substrate fordelivery of at least one compound.

[0026] Other aspects of the present invention comprise compositions andmethods for microarray devices. Such microarray devices and methodscomprise a variety of microarrays that may be used, for example, tostudy and monitor gene expression in response to treatment with thecompounds of the present invention. The microarrays may comprise nucleicacid sequences, carbohydrates or proteins that are determinative forspecific cells, tissues, species, disease states, prognoses, diseaseprogression, or any other combination of molecules that can be used todetermine an effect of one or more of the compounds of the presentinvention. Other embodiments of the present invention comprise methodsusing databases and computer applications.

BRIEF DESCRIPTION OF THE FIGURES

[0027]FIG. 1. ¹H NMR ofN-(3-Chloro-4-methoxy-phenyl)-N′-cyclohexylmethyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0028]FIG. 2. ¹H NMR ofN-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine.

[0029]FIG. 3. ¹H NMR ofN-(3-Chloro-4-methoxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-(1-propyl-butyl)-[1,3,5]triazine-2,4,6-triamine.

[0030]FIG. 4. ¹H NMR ofN-(1-Aza-bicyclo[2.2.2]oct-3-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-)1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4,6-triamine.

[0031]FIG. 5. ¹H NMR ofN2-(3-chloro-4-methoxy-phenyl)-N4-cycloheptyl-N6-methyl-N6-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine.

[0032]FIG. 6. ¹H NMR ofN-Cycloheptyl-N′-ethyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0033]FIG. 7. ¹H NMR ofN-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-pyrrolidin-1-yl-[1,3,5]triazine-2,4-diamine.

[0034]FIG. 8. ¹H NMR ofN-Cyclohexylmethyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine.

[0035]FIG. 9. ¹H NMR of6-Chloro-N-cycloheheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0036]FIG. 10. ¹H NMR of(3-Chloro-4-methoxy-phenyl)-(4,6-dichloro-[1,3,5]triazin-2-yl)-amine.

[0037]FIG. 11. ¹H NMR ofN-(3-Chloro-4-methoxy-phenyl)-N′-isopropyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0038]FIG. 12. ¹H NMR ofN2-(3-chloro-4-methoxy-phenyl)-N4-isopropyl-N6-methyl-N6-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine.

[0039]FIG. 13. ¹H NMR of 5-{4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-pentan-1-ol.

[0040]FIG. 14. ¹H NMR of5-[4-(3-chloro-4-methoxy-phenylamino)-6-(methyl-piperidin-4-yl-amino)-1,3,5-triazin-2-ylamino]-pentan-1-ol.

[0041]FIG. 15. ¹H NMR of6-Chloro-N,N″-bis-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0042]FIG. 16. ¹H NMR ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(4-methyl-cyclohexyl)-[1,3,5]triazine-2,4,6-triamine.

[0043]FIG. 17. ¹H NMR ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine.

[0044]FIG. 18. ¹H NMR ofN-Butyl-N′-(3-chloro-4-methoxy-phenyl)-N″-(1-methyl-piperidin-4-yl)-N-propyl-[1,3,5]triazine-2,4,6-triamine.

[0045]FIG. 19. ¹H NMR ofN2-Butyl-N4-(3-chloro-4-methoxy-phenyl)-N6-methyl-N6-piperidin-4-yl-N2-propyl-1,3,5-triazine-2,4,6-triamine.

[0046]FIG. 20. ¹H NMR of6-Cyclohexylmethoxy-N,N′-bis-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4-diamine.

[0047]FIG. 21. ¹H NMR of(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl)-amine.

[0048]FIG. 22. ¹H NMR ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-1,3,5-triazine-2,4-diamine.

[0049]FIG. 23. ¹H NMR of(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-chloro-4-methoxy-phenyl)-amine.

[0050]FIG. 24. ¹H NMR of6-Cyclohexylmethoxy-N-(1-ethyl-pyrrolidin-2-ylmethyl)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0051]FIG. 25. ¹H NMR ofN-(3-Chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine.

[0052]FIG. 26. ¹H NMR ofN-Azepan-1-yl-N′-(3-chloro-4-methoxy-phenyl)-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0053]FIG. 27. ¹H NMR ofN4-(3-chloro-4-methoxy-phenyl)-N6-methyl-N2-perhydro-azepin-1-yl-N6-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine.

[0054]FIG. 28. ¹H NMR ofN-Azepan-1-yl-6-chloro-N′-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0055]FIG. 29. ¹H NMR ofN″-(3-chloro-4-methoxy-phenyl)-N,N′-bis-perhydro-azepin-1-yl-1,3,5-triazine-2,4,6-triamine.

[0056]FIG. 30. ¹H NMR ofN-(3-Bromo-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0057]FIG. 31. ¹H NMR ofN-(1-benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]-2,4,6-triamine.

[0058]FIG. 32. ¹H NMR of2-chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl-amino]-1,3,5-triazin-2-ylamino}-phenol.

[0059]FIG. 33. ¹H NMR ofN2-cycloheptyl-N4-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine.

[0060]FIG. 34. ¹H NMR ofN2-cycloheptyl-N4-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine.

[0061]FIG. 35. ¹H NMR ofN2-cyclohexylmethyl-N4-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine.

[0062]FIG. 36. ¹H NMR of N2-cyclohexylmethyl-N4-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine.

[0063]FIG. 37. ¹H NMR of({4-cycloheptylamino-6-[((S)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile.

[0064]FIG. 38. ¹H NMR of({4-cycloheptylamino-6-[((R)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile.

[0065]FIG. 39. ¹H NMR ofN2-[(1-ethyl-2-pyrrolidinyl]-N4-(3-fluoro-4-methoxyphenyl)-6-[(S)-2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5-triazine-2,4-diamine.

[0066]FIG. 40. ¹H NMR of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine.

[0067]FIG. 41. ¹H NMR ofN-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0068]FIG. 42. ¹H NMR of4-(3-Chloro-4-methoxy-phenylamino)-6-cycloheptylamino-1,3,5-triazin-2-ol.

[0069]FIG. 43. ¹H NMR ofN2-(3-chloro-4-diethylamino-phenyl)-N4-cycloheptyl-N6-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine.

[0070]FIG. 44. ¹H NMR ofN2-cycloheptyl-N4-(2-dimethylamino-ethyl)-N6-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine.

[0071]FIG. 45. ¹H NMR of({4-cycloheptylamino-6-[1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile.

[0072]FIG. 46. ¹H NMR ofN,N′-di-n-propyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine.

[0073]FIG. 47. ¹H NMR ofN,N′-dicyclopropyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine.

[0074]FIG. 48. ¹H NMR ofN2-Cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine.

[0075]FIG. 49. ¹H NMR ofN2-Cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine.

[0076]FIG. 50. ¹H NMR ofN2-cycloheptyl-N4-(3-fluoro-4-methoxyphenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine,hydrogen chloride salt.

[0077]FIG. 51. ¹H NMR ofN2-(3-chloro-4-diethylamino-phenyl)-N4-cycloheptyl-N6-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamineS42-63hydrogen chloride salt.

[0078]FIG. 52. ¹H NMR ofN2-cycloheptyl-N4-(1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triaminehydrogen chloride salt.

[0079]FIG. 53. Chart showing the effects of compounds in an assay whereglycated human serum albumin (G-HSA) induces IL-6 production.

[0080]FIG. 54. Chart showing the effects of compounds in anantiproliferative assay.

[0081]FIG. 55. ¹H NMR of 4-benzyloxy-3-chloro-phenylamine.

[0082]FIG. 56. ¹H NMR ofN-(4-benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0083]FIG. 57. ¹H NMR ofN-(4-benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0084]FIG. 58. ¹H NMR of4-[4-cycloheptylamino-6-(methyl-piperidin-4-yl-amino)-[1,3,5]triazin-2-ylamino]-phenol.

[0085]FIG. 59. ¹H NMR of4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5-triazin-2-ylamino}-phenol.

[0086]FIG. 60. ¹H NMR of2-Chloro-4-(4,6-dichloro-[1,3,5]triazin-2-ylamino)-phenol.

[0087]FIG. 61. ¹H NMR of2-Chloro-4-(4-chloro-6-cycloheptylamino-[1,3,5]triazin-2-ylamino)-phenol.

[0088]FIG. 62. ¹H NMR of2-Chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-phenol.

[0089]FIG. 63. ¹H NMR ofN-(1-Benzyl-piperidin-4-yl)-N′-(3-fluoro-4-methoxy-phenyl)-N″-cycloheptyl-1,3,5]triazine-2,4,6-diamine.

[0090]FIG. 64. ¹H NMR ofN-(1-Benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-diamine.

[0091]FIG. 65. ¹H NMR ofN-Cycloheptyl-N′-(4-methoxy-phenyl)-N″-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine.

[0092]FIG. 66. ¹H NMR of6-Chloro-N-cyclopropyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0093]FIG. 67. ¹H NMR ofN-Cyclopropyl-N′-(3-flouro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0094]FIG. 68. ¹H NMR ofN-Cyclopropyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine.

[0095]FIG. 69. ¹H NMR of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cyclopropyl-[1,3,5]triazine-2,4-diamine.

[0096]FIG. 70. ¹H NMR ofN-Cyclopropyl-N′-(3-chloro-4-methoxy-phenyl)-N″-methyl-″N-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine.

[0097]FIG. 71. ¹H NMR of6-Chloro-N,N′-bis-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine.

[0098]FIG. 72. ¹H NMR ofN-(1-Ethyl-pyrrolidin-2-ylmethyl)-N′,N″-bis-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine.

[0099]FIG. 73. ¹H NMR of1-[3-{4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazine-2-yloxy}piperidino]-1-ethanone.

[0100]FIG. 74. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine.

[0101]FIG. 75. ¹H NMR of1-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-3-ol.

[0102]FIG. 76. ¹H NMR of1-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-4-ol.

[0103]FIG. 77. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine.

[0104]FIG. 78. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-4-piperidyloxy)-1,3,5-triazine-2,4-diamine.

[0105]FIG. 79. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1,4-thiazinan-4-yl)-1,3,5-triazine-4,2-diamine.

[0106]FIG. 80. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-fluorophenoxy)-1,3,5-triazine-4,2-diamine.

[0107]FIG. 81. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-[2-(2-fluorophenoxy)ethoxy]-1,3,5-triazine-4,2-diamine.

[0108]FIG. 82. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(6-methyl-2-pyrylmethoxy]-1,3,5-triazine-4,2-diamine.

[0109]FIG. 83. ¹H NMR of3-[4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazin-2-yloxy]-2-ethyl-4H-4-pyranone.

[0110]FIG. 84. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-isopropoxy-1,3,5-triazine-2,4-diamine.

[0111]FIG. 85. ¹H NMR of 1-[4-(3-chloro-4-methoxyanilino)-cycloheptylamino-1,3,5-triazin-2-yl]-2-azoloamymethanol.

[0112]FIG. 86. ¹H NMR ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(4-methylpiperzino)-1,3,5-triazine-4,2-diamine.

DETAILED DESCRIPTION OF THE INVENTION

[0113] It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, constructs, and reagentsdescribed herein and as such may vary. It is also to be understood thatthe terminology used herein is for the purpose of describing particularembodiments only, and is not intended to limit the scope of the presentinvention which will be limited only by the appended claims.

[0114] All publications and patents mentioned herein are incorporatedherein by reference for the purpose of describing and disclosing, forexample, the constructs and methodologies that are described in thepublications, which might be used in connection with the presentlydescribed invention. The publications discussed above and throughout thetext are provided solely for their disclosure prior to the filing dateof the present application. Nothing herein is to be construed as anadmission that the inventors are not entitled to antedate suchdisclosure by virtue of prior invention.

[0115] I. Description of Compounds

[0116] In one aspect, the present invention encompasses novel organiccompounds that are generally described as N², N⁴,N⁶-tris(amino)-1,3,5-triazines which are represented by the names inTable 1 and the structural formulas in the remaining Tables 2 andfollowing. Representative compounds of this invention can be describedby the general structural formula below, where N^(A), N^(B) and N^(C)are pendant substituted amino groups attached to 1,3,5-triazines at the2, 4 and 6 positions.

[0117] Thus, the typical compound encompassed by the present inventionincludes triazine compounds comprising the following structure:

[0118] In this typical embodiment, each pendant NR₁R₂, NR₃R₄, and NR₅R₆amino group can represent a primary, secondary, or tertiary amine whenbonded to the triazine core, including a cyclic secondary amidesubstitutent (for example a pyrrolidin-N-yl group), and a range of othersubstituents as described herein. Compositions in accordance with thepresent invention also comprise analogs of the tris(amino) compounds,for example, compounds that are prepared as intermediate compounds inthe synthesis of the tris(amino)triazine compounds indicated above, orcompounds that represent a partially substituted trizaine core. Many ofthe syntheses of triazine compounds of this invention typically usecyanuric chloride C₃N₃Cl₃ as a starting compound, therefore intermediatespecies such as bis(amino)chlorotriazine compounds, or aminodiclorotriazine compounds shown below, where N^(A) and N^(B) are pendantsubstituted amino groups as described above, are also encompassed bythis invention.

[0119] Compositions in accordance with the present invention alsocomprise analogs of the tris(amino)triazine compounds indicated above,including compounds that are isolated as byproducts in the synthesis ofthe tris(amino)triazine compounds, a general formula of which is shownbelow, where E=O or S. An example of such a compound is abis(amino)alkoxy triazine compound.

[0120] In general terms, the compounds and compositions in accordancewith the present invention comprise analogs of the tris(amino)triazinecompounds of the following general structure:

[0121] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;wherein:

[0122] R¹ is in each occurrence independently selected from —H; alkyl,cycloalkyl, alkenyl, cycloalkenyl, cycloalkadienyl, alkynyl, aralkyl,aralkenyl, aralkynyl, heteroalkyl, alkoxy, alkylthio, alkylamino, ordialkylamino, each of which having up to 12 carbon atoms and includinglinear or branched derivatives thereof, cyclic derivatives thereof,substituted derivatives thereof, heteroatom derivatives thereof, orheterocyclic derivatives thereof, aryl; heteroaryl; aryloxy; arylthio;halogen; or amino;

[0123] G is selected from NR¹ or O;

[0124] E is selected from CH or N;

[0125] z is an integer from 0 to 3;

[0126] X¹ is selected from R¹, NR¹ ₃ ⁺, CN, NO₂, CO₂R¹, C(O)NR¹ ₂,CH═CR¹ ₂, C≡CR¹, C(O)R¹, SO₂R¹, SO₂OR¹, or NC(O)R¹, or X¹ and X²together is a fused aryl, pyridine, dioxane, pyrrole, pyrrolidine,furan, or thiophene ring; with the proviso that the R¹ moiety of theC(O)R¹ substituent in the X¹ position excludes amino or dialkylaminowhen X¹ is C(O)R¹;

[0127] X² is selected from R¹; CX_(x)H_(3-x), wherein X is a halogen andx is an integer from 0 to 3; OR¹; SR¹; NR¹ ₂; CN; C(O)OR¹; NC(O)R¹;4-morpholinyl; 4-methyl-1-piperazinyl; OR², wherein R² is selected fromCH₂OCH₃, CH₂OCH₂OCH₃, CH₂OCH₂CH₂OCH₃, CH₂SCH₃, or C(O)R¹; SR³, whereinR³ is selected from CH₂OCH₃, CH₂OCH₂CH₂OCH₃, CH₂OCH₂CH(CH₃)₂,CH₂NHC(O)CH₃, or SR¹; OM or SM, wherein M is selected from Li, Na, K,Mg, or Ca;

[0128] AY¹ is halogen, or A is selected from NR¹ or O, and Y¹ isselected from R¹; CR⁴ ₃; NR⁴ ₂; OR⁴; or SR⁴;

[0129] wherein n is an integer from 0 to 8, m is an integer from 1 to 8,Z¹ is independently selected from CR¹ or N, Z² is independently selectedfrom CR¹ ₂, NR¹, O, or S, with the proviso that two O or S atoms are notlocated adjacent to each other, and the proviso that no more than two Z²moieties are NR¹;

[0130] R⁴ is in each occurrence independently selected from linear orbranched alkyl, cycloalkyl, cycloalkenyl, cycloalkadienyl, alkenyl,alkynyl, aralkyl, aralkenyl, aralkynyl, heteroalkyl, alkoxy, alkylthio,alkylamino, or dialkylamino, each of which having up to 10 carbon atoms,—H, aryl, heteroaryl, aryloxy, arylthio, halogen, amino, NR¹₂-substituted derivatives thereof, OR¹-substituted derivatives thereof,SR¹-substituted derivatives thereof, or halogen-substituted derivativesthereof; and

[0131] DY² is halogen, or D is selected from NR¹ or O wherein R¹ isdefined as above, and

[0132] Y² is selected from R¹,

[0133] wherein Z¹ is independently selected from N or CR⁴ and Z² isindependently selected as defined above, with the proviso that two O orS atoms are not located adjacent to each other, and with the provisothat no more than two Z² moieties are NR¹. The compounds of the presentinvention according to this general description do not include thosethat encompass the unique combination of substituents that would providethe following compounds:

[0134]N-Cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl-[1,3,5]triazine-2,4,6-triamine;

[0135]N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine;

[0136][4-(4-Benzyl-piperazin-1-yl)-6-morpholin-4-yl-[1,3,5]triazin-2-yl]-(4-methoxy-phenyl)-amine;

[0137]N-Cycloheptyl-6-morpholin-4-yl-N′-naphthalen-2-yl-[1,3,5]triazine-2,4-diamine;

[0138] N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-morpholin-4-yl-[1,3,5]triazine-2,4-diamine;

[0139]N-Cycloheptyl-6-morpholin-4-yl-N′-phenyl-[1,3,5]triazine-2,4-diamine;

[0140]N-Cycloheptyl-N′-(4-methoxy-phenyl)-6-morpholin-4-yl-[1,3,5]triazine-2,4-diamine;

[0141]N-Benzyl-N′-cycloheptyl-N″-(4-methoxy-phenyl)-N-methyl-[1,3,5]triazine-2,4,6-triamine;

[0142]N-(2-[1,3]Dioxolan-2-yl-ethyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl-[1,3,5]triazine-2,4,6-triamine;or

[0143]N-Cyclopropyl-N′-methyl-N′(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl-[1,3,5]triazine-2,4,6-triamine.

[0144] Because this invention encompass compounds that representsaturated derivatives of the above general structure, and compounds thatinclude various states of unsaturation (for example, -ene, -diene,-triene, and -yne derivatives of the above compounds), then the aryl orpyridyl ring shown in the general formula above can be partially orcompleted hydrogenated in this invention. As a result, the C₅E ring inthe above structure can represent a cylcohexyl or piperidynl ring thatis X¹ and X² substituted. In general terms, X¹ usually, but not always,represent an electron withdrawing group such as halide or nitro, whileX² usually, but not always, represents an electron donating group suchas alkoxide or amino.

[0145] As indicated in the general structure above, AY¹ and DY²typically represent an NR¹ moiety (where R¹ is defined above), in whichcase these substituents constitute a portion of amino or substitutedamino group and therefore, the compound itself constitutes a triazine.In this case, Y¹ and Y² may be selected from a wide range ofsubstituents, including, but not limited to, cycloalkyl with up to 10carbon atoms;

[0146] wherein n is from 1 or 2;

[0147] ;linear or branched alkyl with up to 10 carbon atoms; CH₂R¹;(CHR¹)_(x)NR¹ ₂ wherein x is 1-6; CH₂R¹; (CHR¹)_(x)OR¹ wherein x is from1 to 6;

[0148] wherein x is from 3 to 5;

[0149] CH₂CF₃; (CHR¹)_(x)Z¹ wherein x is from 1 to 6 and Z¹ is selectedfrom NR¹ ₂,

[0150] wherein y is from 3 to 5,

[0151] In these examples, R¹ is independently selected as defined above.These are merely representative examples of the definitions of the Y¹and Y² substitutents, and are not intended to be exclusive.

[0152] It is also noted that AY¹ together, and DY² together, can alsorepresent a wide range of chemical moieties bonded to the triazine coresuch as halide or secondary amino groups such as

[0153] In the latter cases, the amino substituent groups are termedsecondary amino groups, however upon bonding to the triazine core, theamino nitrogens become tertiary amine moieties. Examaples of AY¹together and DY² together include, limited to: halide;

[0154] wherein x is from 3 to 5;

[0155] wherein x is from 0 to 6;

[0156] wherein Z² is selected from R¹,

[0157] C(O)R¹, C(O)OR¹, pyridinyl, aryl,

[0158] wherein x is from 0 to 6, and wherein R¹ is independentlyselected as defined above. These are also merely representative examplesof the definitions of these substitutents, and are not intended to beexclusive.

[0159] Representative compounds in accordance with the present inventionare presented in Table 1. This table is not intended to be exclusive ofthe compounds of the present invention, but rather exemplary of thetriazine compounds that are encompassed by this invention. TABLE 1COMPOUND NUMBER COMPOUND NAME 1N²-(4-bromo-1-naphthyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 2N²-(4-chloro-1-naphthyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 3N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-quinolinyl)-1,3,5-triazine-2,4,6-triamine 4N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(6-quinolinyl)-1,3,5-triazine-2,4,6-triamine 5N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(8-quinolinyl)-1,3,5-triazine-2,4,6-triamine 6N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-[1-(2-naphthyl)ethyl]-1,3,5-triazine-2,4,6-triamine 7N²-cycloheptyl-N⁴-(3,4-dichlorophenyl)-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 8N²-cycloheptyl-N⁴-(3,4-difluorophenyl)-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 9N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-[4-(trifluoromethoxy)phenyl]-1,3,5-triazine-2,4,6-triamine 10N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-fluorophenyl)-1,3,5-triazine-2,4,6-triamine 114-[(4-cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-amino]benzonitrile 12N²-(4-chlorophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 13N²-(4-bromophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 14 Ethyl4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-amino]benzoate 15N²-(1,1′-biphenyl-4-yl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 16N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluorophenyl)-1,3,5-triazine-2,4,6-triamine 17N²-(3-chlorophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 18N²-(3-bromophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 19 Ethyl3-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-amino]benzoate 20N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(2-fluorophenyl)-1,3,5-triazine-2,4,6-triamine 21N²-(2-chlorophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 22N²-(2-bromophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 23N²-(1,3-benzodioxol-5-yl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 24N²-cycloheptyl-N⁴-(2,3-dihydro-1,4-benzodioxin-6-yl)-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 25N²-cycloheptyl-N⁴-[4-(dimethylamino)phenyl]-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 26N²-[3-chloro-4-(diethylamino)phenyl]-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 27N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-[4-(4-morpholinyl)phenyl]-1,3,5-triazine-2,4,6-triamine 28N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-[4-(4-methyl-1-piperazinyl)phenyl]-1,3,5-triazine-2,4,6-triamine 29N-{4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]phenyl}acetamide30 N-{3-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]phenyl}acetamide31 N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 32N²-cycloheptyl-N⁴-(4-ethoxyphenyl)-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 33N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-[4-(methylthio)phenyl]-1,3,5-triazine-2,4,6-triamine 34N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(2-pyridinyl)-1,3,5-triazine-2,4,6-triamine 35N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(2-methylphenyl)-1,3,5-triazine-2,4,6-triamine 36N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-phenoxyphenyl)-1,3,5-triazine-2,4,6-triamine 37N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-methylphenyl)-1,3,5-triazine-2,4,6-triamine 38N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-methylphenyl)-1,3,5-triazine-2,4,6-triamine 392-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-amino]-4-methyl-3-thiophenecarboxamide 40N²-(4-chlorophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-N²-methyl-1,3,5-triazine-2,4,6-triamine 413-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-(phenyl)amino]propanenitrile 42N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-methoxyphenyl)-N⁶-methyl-1,3,5-triazine-2,4,6-triamine 43N²-cycloheptyl-N⁴-(2,4-difluorophenyl)-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-methyl-1,3,5-triazine-2,4,6-triamine 44[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)(phenyl)amino]acetonitrile 45N²-(3-chlorophenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-N²-methyl-1,3,5-triazine-2,4,6-triamine 46N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-methyl-N⁶-[2-(trifluoromethyl)phenyl]-1,3,5-triazine-2,4,6-triamine 47N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-methyl-N⁶-[4-(trifluoromethoxy)phenyl]-1,3,5-triazine-2,4,6-triamine 48N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-N⁶-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine 49N-benzoyl-4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]benzenesulfonamide50 N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(2-naphthyl)-1,3,5-triazine-2,4,6-triamine 51N²-ethyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 52N²-(tert-butyl)-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 53N²-benzyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 54N²-cyclooctyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 55N²-cyclohexyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 56N²-cyclopentyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 57N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-(1-pyrrolidinyl)-1,3,5-triazine-2,4-diamine 58N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-hexahydro-1H-azepin-1-yl-1,3,5-triazine-2,4-diamine 59N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-octahydro-1(2H)-quinolinyl-1,3,5-triazine-2,4-diamine 60N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(4-methylcyclohexyl)-1,3,5-triazine-2,4,6-triamine 61N²-(1-ethyl-pyrrolidin-2-ylmethyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-((S)-2-methoxymethyl-pyrrolidin-1-yl)-1,3,5-triazine-2,4-diamine 62N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5-triazine-2,4-diamine 636-(4-acetyl-1-piperazinyl)-N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine 64 Ethyl4-{4-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-N⁶-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2-yl}-1-piperazinecarboxylate 65N²-(cyclohexylmethyl)-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 66N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(2-furylmethyl)-1,3,5-triazine-2,4,6-triamine 67N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(2,2,2-trifluoroethyl)-1,3,5-triazine-2,4,6-triamine 68N²-[2-(dimethylamino)ethyl]-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 69N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-{4-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1-piperazinyl}-1,3,5-triazine-2,4-diamine 70 N²,N⁴-bis[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 71N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[2-(1-piperidinyl)ethyl]-1,3,5-triazine-2,4,6-triamine 72N⁶-[4-(1,3-benzodioxol-5-ylmethyl)-1-piperazinyl]-N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine 73N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[4-(2-pyridinyl)-1-piperazinyl]-1,3,5-triazine-2,4-diamine 741-[3-({4-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2-yl}amino)propyl]-2- pyrrolidinone75 N²-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[3-(1H-imidazol-1-yl)propyl]-1,3,5-triazine-2,4,6-triamine 76N²-cycloheptyl-N⁴-ethyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 77N²-(tert-butyl)-N⁴-cycloheptyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 78N²-benzyl-N⁴-cycloheptyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 79N²-cycloheptyl-N⁴-cyclooctyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 80N²-cycloheptyl-N⁴-cyclohexyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 81N²-cycloheptyl-N⁴-cyclopentyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 82N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-(1-pyrrolidinyl)-1,3,5-triazine-2,4-diamine 83N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-hexahydro-1H-azepin-1-yl-1,3,5-triazine-2,4-diamine 84N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-octahydro-1(2H)-quinolinyl-1,3,5-triazine-2,4-diamine 85N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(4-methylcyclohexyl)-1,3,5-triazine-2,4,6-triamine 86N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-[(2S)-2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5-triazine-2,4-diamine 87N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5-triazine-2,4-diamine 886-(4-acetyl-1-piperazinyl)-N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine 89ethyl-4-{4-(cycloheptylamino)-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2-yl}-1-piperazinecarboxylate 90N²-cycloheptyl-N⁴-(cyclohexylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 91N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(2-furanylmethyl)-1,3,5-triazine-2,4,6-triamine 92N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(2,2,2-trifluoroethyl)-1,3,5-triazine-2,4,6-triamine 93N²-cycloheptyl-N⁴-[2-(dimethylamino)ethyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 94N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-{4-[2-oxo-(1-pyrrolidinyl)ethyl]-1-piperazinyl}-1,3,5-triazine-2,4-diamine 95N²-cycloheptyl-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 96N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[2-(1-piperidinyl)ethyl]-1,3,5-triazine-2,4,6-triamine 976-[4-(1,3-benzodioxol-5-ylmethyl)1-piperazinyl]-N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine 98N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-6-[4-(2-pyridinyl)-1-piperazinyl]-1,3,5-triazine-2,4-triamine 991-[3-({4-(cycloheptylamino)-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2-yl}amino)propyl]-2-pyrrolidinone 100N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[3-(1H-imidazol-1-yl)propyl]-1,3,5-triazine-2,4,6-triamine 101(3-Chloro-4-methoxy-phenyl)-(4,6-dichloro-[1,3,5]triazin-2-yl)-amine 1026-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cyclohexylmethyl-[1,3,5]triazine-2,4-diamine 103N-(3-Chloro-4-methoxy-phenyl)-N′-cyclohexylmethyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 1046-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-(1-propyl-butyl)-[1,3,5]triazine-2,4-diamine 105N-(3-Chloro-4-methoxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-(1-propyl-butyl)-[1,3,5]triazine-2,4,6-triamine 106N-(3-Chloro-4-methoxy-phenyl)-N′-isopropyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 107N²-(3-chloro-4-methoxy-phenyl)-N⁴-isopropyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 1085-{4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-pentan-1-ol 1095-[4-(3-chloro-4-methoxy-phenylamino)-6-(methyl-piperidin-4-yl-amino)-1,3,5-triazin-2-ylamino]-pentan-1-ol 110N-Butyl-6-chloro-N′-(3-chloro-4-methoxy-phenyl)-N-propyl-[1,3,5]triazine-2,4-diamine 111N-Butyl-N′-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-N-propyl-[1,3,5]triazine-2,4,6-triamine 112N²-Butyl-N⁴-(3-chloro-4-methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-N²-propyl-1,3,5-triazine-2,4,6-triamine 1132,4-Dichloro-6-cyclohexylmethoxy-[1,3,5]triazine 114(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl)-amine 1156-Cyclohexylmethoxy-N,N′-bis-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4-diamine 1166-Cyclohexylmethoxy-N-(1-ethyl-pyrrolidin-2-ylmethyl)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 117(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-chloro-4-methoxy-phenyl)-amine 118N,N′-Bis-(3-chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-1,3,5-triazine-2,4-diamine 119N-(3-Chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine 1206-Chloro-N,N″-bis-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 121 N,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 122N,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine 123N-(3-Bromo-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 124(4,6-Dichloro-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl)-amine 1256-Chloro-N-cyclohexylmethyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 126N-Cyclohexylmethyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine 1276-Chloro-N-cycloheheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 128N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-pyrrolidin-1-yl-[1,3,5]triazine-2,4-diamine 129N-Cycloheptyl-N′-ethyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 130N-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine 1312-[4-chloro-6-(3-chloro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-propane-1,3-diol 1322-{4-(3-chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-propane-1,3-diol 1336-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine 134N-(1-benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]-2,4,6-triamine 135N²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 136N²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine 137N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 1382-chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl-amino]-1,3,5-triazin-2-ylamino}-phenol 139N²-cycloheptyl-N⁴-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 140N²-cycloheptyl-N⁴-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 141N²-cyclohexylmethyl-N⁴-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine 142N²-cyclohexylmethyl-N⁴-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine (42, Scheme 23)143 ({4-cycloheptylamino-6-[((S)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile (43, Scheme 24) 144({4-cycloheptylamino-6-[((R)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile 145N²-[(1-ethyl-2-pyrrolidinyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-[(S)-2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5-triazine-2,4-diamine 146N²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 1474-(3-Chloro-4-methoxy-phenylamino)-6-cycloheptylamino-1,3,5-triazin-2-ol 148N-(1-Aza-bicyclo[2.2.2]oct-3-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-)1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4,6-triamine 149N²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine 150N²-cycloheptyl-N⁴-(2-dimethylamino-ethyl)-N⁶-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine 151({4-cycloheptylamino-6-[1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile 152N-Azepan-1-yl-6-chloro-N′-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine 153N″-(3-chloro-4-methoxy-phenyl)-N,N′-bis-perhydro-azepin-1-yl-1,3,5-triazine-2,4,6-triamine 154N-Azepan-1-yl-N′-(3-chloro-4-methoxy-phenyl)-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine 155N⁴-(3-chloro-4-methoxy-phenyl)-N⁶-methyl-N²-perhydro-azepin-1-yl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 156N,N′-di-n-propyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine 157N,N′-dicyclopropyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine 158N²-Cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine 159N²-Cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 160N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine, hydrogen chloridesalt 161 [N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]trizaine-2,4,6-triamine, hydrogen chloridesalt 162 N²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine 163N²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine hydrogen chloridesalt 164N²-cycloheptyl-N⁴-(1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine hydrogen chloride salt 165N²-(cyclohexylmethyl)-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-fluoro-3-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine hydrogen chloridesalt 166({4-cycloheptylamino-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile hydrogen chloride salt 167N²-cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine maleate salt 168N²-cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine citrate salt 169N²-cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine succinate salt 170N-(3-Bromo-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine hydrogen chloridesalt

[0160] In general terms, the compositions in accordance with the presentinvention also comprise tris(amino)triazine compounds of the followingstructure:

[0161] wherein R₁ to R₆ represent H, alkyl, aryl, alkenyl, alkynyl,aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl,heteroaryl, halide, alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy,amino, alkylamino, dialkylamino and the like, including straight orbranched chain derivatives thereof, cyclic derivatives thereof,substituted derivatives thereof, heteroatom derivatives thereof,heterocyclic derivatives thereof, functionalized derivatives thereof,salts thereof, isomers thereof, or combinations thereof.

[0162] For example, a typical substituent R₁ to R₆ is a substitutedalkyl, in which the substituent is a heterocyclic derivative. Examplesof nitrogen-containing heterocyclic moieties include, but are notlimited to groups such as pyridinyl (derived from pyridine, bondedthrough a ring carbon), piperidinyl (derived from piperidine and bondedthrough the ring nitrogen atom or a ring carbon), and pyrrolidinyl(derived from pyrrolidine and bonded through the ring nitrogen atom or aring carbon).

[0163] Examples of substituted or functionalized derivatives of R₁ to R₆include, but are not limited to, moieties containing substituents suchas acyl, formyl, hydroxy, acyl halide, amide, amino, azido, acid,alkoxy, aryloxy, halide, carbonyl, ether, ester, thioether, thioester,nitrile, alkylthio, arylthio, sulfonic acid and salts thereof, thiol,alkenyl, alkynyl, nitro, imine, imide, alkyl, aryl, combinationsthereof, and the like. Moreover, in the case of alkylated derivatives ofthe recited moieties, the alkyl substitutent may be pendant to therecited chemical moiety, or used for bonding to the amine nitrogenthrough the alkyl substituent.

[0164] Examples of chemical moieties R₁ to R₆ of the present inventionfurther include, but are not limited to: H; methyl; ethyl; propyl;butyl; pentyl; hexyl; heptyl; octyl; ethenyl; propenyl; butenyl;ethynyl; propynyl; butynyl; cyclobutyl; cyclopentyl; cyclohexyl;cyclobutenyl; cyclopentenyl; cyclohexenyl; phenyl; tolyl; xylyl; benzyl;naphthyl; pyridinyl; furanyl; tetrahydro-1-napthyl; piperidinyl;indolyl; indolinyl; pyrrolidinyl; 2-(methoxymethyl)pyrrolidinyl;piperazinyl; quinolinyl; quinolyl; alkylated-1,3-dioxolane; triazinyl;morpholinyl; phenyl pyrazolyl; indanyl; indonyl pyrazolyl; thiadiazolyl;rhodaninyl; thiolactonyl; dibenzofuranyl; benzothiazolyl;homopiperidinyl; thiazolyl; quinonuclidinyl; isoxazolidinonyl; anyisomers, derivatives, or substituted analogs thereof; or any substitutedor unsubstituted chemical groups such as alcohol, ether, thiol,thioether, tertiary amine, secondary amine, primary amine, ester,thioester, carboxylic acid, diol, diester, acrylic acid, acrylic ester,methionine ethyl ester, benzyl-1-cysteine ethyl ester, imine, aldehyde,ketone, amide, or diene.

[0165] Further examples of chemical moieties R₁ to R₆ of this inventioninclude, but are not limited to, the following species or substituted oralkylated derivatives of the following species, covalently bonded to theamine nitrogen: furan; tetrahydrofuran; indole; piperazine; pyrrolidine;pyrrolidinone; pyridine; quinoline; anthracene; tetrahydroquinoline;naphthalene; pyrazole; imidazole; thiophene; pyrrolidine; morpholine;and the like. One feature of the recited species or substituted oralkylated derivatives of these species, is that they may be covalentlybonded to the amine nitrogen in any fashion, including through thependant substituent or alkyl group, through the heteroatom asappropriate, or through a ring atom as appropriate, as understood by oneof ordinary skill in the art.

[0166] The chemical moieties R₁ to R₆ of the present invention alsoinclude, but are not limited to, cyclic alkanes and alkenes and includebridged and non-bridged rings. Examples of bridged rings include, butare not limited to, groups such as norbornyl; norbonadienyl, adamantyl;6-azabicyclo[3.2.1]octanyl; 3-azabicyclo[2.2.2]octanyl, and the like.

[0167] In one embodiment of the present invention, NR₁R₂, NR₃R₄, orNR₅R₆ are derived from a cyclic secondary amine. Examples of a cyclicamino chemical moiety of the present invention include, but are notlimited to piperidine; 4-benzyl-piperidine; 3-piperidinemethanol;moropholine; 4-piperidinopiperidine; 1-(2-amino-methyl)-piperazine;decahydroquinoline; 1,2,3,4-tetrahydro-pyridoindole (either aminemoiety); 3-amino-5-phenyl pyrazole; 3-aminopyrazole; bistidinol;hexamethyleimine; 4-hydroxypiperidine; 2-piperidinemethanol;1,3,3-trimethyl-6-azabicyclo[3.2.1]octane; 3-pyrrolidinol;1-methylpiperazine; 2-ethyl-piperidine; 1,2,3,4-tetrahydroisoquinoline;3-aminopyrrolidine; 2,6-dimethylmorpholine;2,3,4-tetrahydroisoquinoline; 1,2,3,4-tetrahydroquinoline;1-(2-methoxyphenyl) piperazine; 2,6-dimethylpiperazine (either aminemoiety); iminodibenzyl; 5-methoxytryptamine; 4,4′-bipiperidine;1-(2-hydroxyethyl)piperazine; 4-methylpiperidine; and the like.

[0168] Importantly, the general structure of the present inventionencompasses all states of saturation of the substitutents shown, such asall ene, diene, triene, and yne derivatives of any substitutent. Thegeneral structure also encompasses all conformational isomers,regioisomers, and stereoisomers that may arise from a particular set ofsubstitutents. The general structure also encompasses all enantiomers,diastereomers, and other optical isomers whether in enantiomeric orracemic forms, or mixtures of stereoisomers.

[0169] Preparation of the Focused Library of Compounds

[0170] Many of the compounds of this invention were prepared in aparallel synthetic procedure according to the methods described below.Examples of compounds prepared by the parallel synthesis techniques areprovided in Table 2. These preparations involve reacting the individualamine compounds (monomers) with cyanuric chloride, which are alsopresented in Table 2, along with the chemical structures of compoundsprepared by the parallel synthesis methods.

[0171] A library of compounds was synthesized according to the presentinvention to afford substituted N², N⁴, N⁶-tris(amino)-1,3,5-triazines,as follows. The design of the compound library was based primarily onstructure 95 shown below. That is, the design of the N², N⁴,N⁶-tris(amino)triazines was focused so that only one of the pendantamino groups (N^(A), N^(B), or N^(C) in the structure above) was changedduring each synthesis, while the other two groups were held constant.The combination of the specific amines employed produced a library ofcompounds of novel composition. Initially, the library was developedusing methyl-(1-methyl-piperidin-4-yl)-amine, holding the cycloheptyland m-fluoroanisidyl groups constant (in structure 95 below). Thesynthesis of the triazines around methyl-(1-methyl-piperidin-4-yl)-aminewas not optimized, and the amine was subsequently replaced with(1-ethyl-pyrrolidin-2-yl)-methylamine which provided a more tractablesynthesis.

[0172] The library of N², N⁴, N⁶-tris(amino)-1,3,5-triazines wasprepared based on the strategy of changing only one pendant amino groupper synthesis, and based on the parent structure 95 shown above. Thelibrary was divided into three subgroups: Libraries I, II, and III(shown in Table 2). Library I includes compounds having unchanged N^(B)and N^(C) groups but different N^(A) groups (6). The pendant amino groupN^(A) was changed according to the specific examples listed below.Library II includes compounds having unchanged N^(A) and N^(C) groupsand different N^(B) groups (7). Pendant amino group N^(B) was changedaccording to the specific examples listed below. Library III includescompounds having unchanged N^(A) and N^(B) groups and different N^(C)groups (8). The pendant amino group N^(C) was changed according to thespecific examples listed below.

[0173] The N², N⁴, N⁶-tris(amino)-1,3,5-triazine compound structuresthat are presented in Tables 2 and following were generated usingISIS-Draw™ version 2.4.0.20, and were generated with the option todisplay unspecified hydrogen atoms if not shown, however, not allhydrogen atoms were displayed in the structures shown. In all structurespresented in any text, table, scheme or figure herein, any hydrogenatoms that are required for any atom to attain its usual valence,whether a carbon atom or a heteroatom, should be inferred if it is notspecifically indicated in a structure.

[0174] One method of preparation of the compounds is shown in the schemebelow. The compounds were prepared by reacting cyanuric chloridesequentially with monomers of primary or secondary amines to afford thedesired 1,3,5-triazine derivatives [1,2,3,4]. Thus, the amine startingcompounds that are used to react with cyanuric chloride are termed“monomers.” The N², N⁴, N⁶-tris(amino-substituted)-1,3,5-triazines wereprepared without the need for purification between each step of thesynthesis, and the final product was isolated by standard procedures.Purification was accomplished using flash column chromatography asneeded. It is within the skill of the art of organic synthesis toprepare, isolate and purify these organic compounds described herein,and to modify the syntheses shown. For example, it is possible tosynthesize the compounds of the present invention by using an excess ofany monomers of primary or secondary amines in any of the three stepsshown in Scheme 1, such that the excess monomer, serves as bothsubstituent for the triazine core, as well as a base, in which casei-Pr₂NEt base can be excluded.

[0175] The pendant amino groups can be substituted by functional groupsdepicted as R₁ to R₆ groups in Scheme 1. The degree of functionality ofa pendant amino group is determined by the structure and complexity ofthe amine monomer, and will affect the overall molecular diversity ofthe N²,N⁴,N⁶-tris(amino-substituted)-1,3,5-triazines. A wide range ofamine monomers may be used in this invention. Once bonded to thetriazine core, the pendant amino groups can be described as secondary ortertiary substituted, depending on the degree of substitution at thenitrogen atom.

[0176] Table 2 presents charts of N²,N⁴, N⁶-tris(amino)-1,3,5-triazinecompounds of Libraries I-III of this invention, respectively, along withthe amine precursor monomers used in the preparation of the compounds.General procedures and synthetic procedures are detailed in Example 1-5.The sequence in which each monomer is added in Scheme 1 is alsopresented in Table 2, where Monomer 1 is added first, Monomer 2 addedsecond, and Monomer 3 is added third. While not intending to be bound bythe following statement, it is believed that this order of addition issignificant, because each synthetic stop necessarily involves thereaction of a monomer with a different triazine precursor. That is,monomer 1 reacts with cyanuric chloride, monomer 2 reacts with an aminodichloro(triazine), and monomer 3 reacts with a diaminochloro(triazine), as shown in Scheme 1. Thus, the order in which themonomers are employed is based on the general synthetic principle thatthe relative nucleophilicity and/or basicity of monomers 1-3 used in thesynthetic scheme should generally increase from monomer 1 to monomer 3.This strategy permits the most nucleophilic and/or basic amine monomerto be reacted with the more sterically congested and presumablyless-reactive diamino chloro(triazine), where its greater reactivity mayassist the reaction proceeding to completion. In some cases, more thanone order of monomer addition will provide the desired product, but thereaction sequences provided in Table 2 represents the optimum syntheticmethods presently known.

[0177] Note that only in a general sense do the substituents indicatedas N^(A), N^(B), and N^(C) in the general structures above correspondwith the actual N², N⁴, N⁶-nomenclature of the N², N⁴,N⁶-tris(amino)triazines. Because the order in which N², A⁴, and N⁶substituents are assigned a 2-, 4-, or 6-position on the triazine coreis dependent on the name of each amino group in the molecule, it is notalways true that one particular amino group always appears as an N², N⁴,or N⁶ substituent, even when only a single substituent is being permutedat one position. For example, many of the compounds of Table 2 containboth cycloheptyl amino and 3-fluoro-4-methoxyphenyl amino groups, yetthese groups take on different 2-, 4-, or 6-positions as a function ofthe name of the third substituent on the triazine core. As a result, thesyntheses are discussed in terms of permuting amino groups at onependant N^(A), N^(B), or N^(C) position (rather than N², N⁴, or N⁶position) in the structure above, while maintaining the other aminogroups constant. Further, note that the compound names used in theTables, Claims and specification were typically generated usingBeilstein's Autonom™ 4.01.188, as well as the earlier CD “stand-alone”version of Beilstein's Autonom™, Autonom 2000. Typically, the compoundnames generated in this fashion were used, regardless of whether thecompound name is an IUPAC, CAS, Beilstein, or other nomenclature. Ineach case however, the names unambiguously identify the compoundspecified.

[0178] A. Amino Groups Derived from Monomer 1

[0179] The sequence of monomer reaction with the triazine core, shown inScheme 1, is Monomer 1, Monomer 2, and Monomer 3, added in that order.Thus, an amino dichloro(triazine) is formed from Monomer 1 and cyanuricchloride. For the first pendant amino group derived from Monomer I andcyanuric chloride, the Monomer 1 amine used and proposed includedprimarily, but not always, aryl amines, specifically phenyl, naphthyl,naphthylalkyl, quinolinyl, heteroaryl derivatives, and the like.

[0180] Specific examples of Monomer 1 used to produce the first pendantamino group in N², N⁴, N⁶-tris(amino-substituted)-1,3,5-triazines, andtheir [Chemical Abstract Registry numbers] include, but are not limitedto, 4-chloroaniline [106-47-9], 3,4-ethylenedioxaniline [22013-33-8],4-bromoanline [106-40-1], ethyl 4-aminobenzoate [94-09-7],4-fluoro-aniline [371-40-4], 4-aminobiphenyl [92-67-1], 3-fluoroaniline[372-19-0], 2-aminonaphthalene [91-59-8], 3-chloroaniline [108-42-9],4-morpholinoaniline [2524-67-6], 3-bromoaniline [591-19-5],4′-aminoacetanilide [122-80-5], ethyl 3-aminobenzoate [582-33-2]m-aminoacetanilide [102-28-3], 2-fluoroaniline [348-54-9]m-anisidine[536-90-3], 2-chloroaniline [[95-51-2], p-phenetidine [156-43-4],2-bromoaniline [615-36-1], 4-(methylthio)aniline [104-96-1],3,4-(methylendioxy) aniline [14268-66-7], 2-aminopyridine [504-29-0],o-toluidine [95-53-4], 2,4-difluoro-N-methylaniline [138564-16-6],4-phenoxyaniline [139-59-3], N-phenylglycinonitrile [3009-97-0],m-toluidine [108-44-1], 3-chloro-N-methylaniline [7006-52-2],p-toluidine 106-49-0], 2-(methylamino)benzotrifluoride,4-chloro-N-methylaniline [932-96-7], 4-aminobenzonitrile [873-74-5],3-anilinopropionitrile, [1075-76-9], tetracaine [94-24-6],N-methyl-p-anisidine [5961-59-1], 3-chloro-p-anisidine [5345-54-0],sulfabenzamide [127-71-9], 3-aminoquinoline [580-17-6], 1-amino-4-bromonaphthalene [2298-07-9], 6-aminoquinoline [580-15-4]1-amino-4-chloronaphthalene, [4684-12-2] 8-aminoquinoline [578-66-5],S-(−)-1-(2-naphthyl)-ethylamine [3082-62-0], 3,4-dichloroaniline[95-76-1], 3,4-difluoroaniline [3863 - 11-4],N-methyl-4-(trifluoromethoxy)aniline [41419-59-4],4-(trifluoromethoxy)aniline [461-82-5],2-amino-4-methylthiophene-3-carboxamide [4651-97-2],N,N-diethyl-N′-phenethylenediamine[1665-59-4],1-(4-amino-phenyl)-4-methylpiperazine hydrochloride[94520-33-9], 2-chloro-N′,N′-diethyl-1,4-phenylenediaminemonohydrochloride [196938-07-5] 2-(dimethylamino)ethyl 4-aminobenzoate[11012-47-2], N,N-dimethyl-1,4-phenylenediamine [1665-95-4].

[0181] B. Amino Groups Derived from Monomer 2

[0182] The reaction of Monomer 2 with a preformed aminodichloro(triazine) provides an intermediate diamino chloro(triazine) inthe synthesis of N², N⁴, N⁶-tris(amino-substituted)-1,3,5-triazines.Thus, for bonding the second pendant amino group to the triazine core,the Monomer 2 amine used and proposed included amines, specificallyalkyl (C₁-C₁₂, straight chain or branched), cycloalkyl (C₃-C₁₀ ringsize), azacyclo (C₂-C₁₀), and benzyl amine derivatives. The ring of thecycloalkyl and azacycloamine, and phenyl ring of the benzyl derivativescan be optionally substituted with one or more moieties, or acombination of moieties, such as, alkyl, alkenyl, alkynyl, phenyl,benzyl, halo, cyano, nitro, hydroxy, thioxy, alkoxy, aryloxy,baloalkyloxy, alkylthio, arylthio, amino, alkyl amino, aryl amino, acyl,carboxyl, amido, sulfonamido, sulfonyl, sulfate, sulfonic acid,morpholino, thiomorpholino, piperazinyl, pyridyl, thienyl, furanyl,pyrroyl, pyrazoyl, phosphate, phosphonic acid, phosphonate and the like.These groups can be represented in protected or unprotected forms usedin standard organic synthesis.

[0183] In addition, any monomer described that has a stereocenterincludes its enantiomers, diastereomers, and optical isomers whether inenantiomeric or racemic forms, or mixtures of stereoisomers. This is toinclude all of the 1,3,5-triazine derivatives and their stereoisomerspresented herein that are formed as a result of using optically active,scalemic or racemic monomers.

[0184] Specific examples of Monomer 2 used to attach the second pendantamino group in the synthesis of N², N⁴,N⁶-tris(amino-substituted)-1,3,5-triazines, and their corresponding[Chemical Abstract Registry numbers] include, but are not limited to,ethylamine [75-04-07], cyclohexanemethylamine [3128-02-8]tert-butylamine [75-64-9], furfurylamine [617-89-0], benzylamine[100-46-9], 2,2,2-trifluroethylamine [753-90-2], cyclooctylamine[5452-37-9N,N-dimethylethylenediamine cyclohexylamine [108-91-8],cyclopentylamine [1003-03-8], 1-(2-aminoethyl)-piperidine [26116-12-1],1-acetylpiperazine [13096-96-3], pyrrolidine [123-75-1],1-piperonylpiperazine [32231-06-4], hexametbyleneimine [111-49-9],1-(2-pyridyl)piperazine [34803-66-2], decahydroquinoline (cis/trans)[2051-28-7], 1-methylpiperazine [109-01-3], 1-(3-aminopropyl)-imidazole[5036-48-6], ethyl 1-piperazine carboxylate [120-43-4],4-methylcyclohexylamine (cis/trans) [6321-23-9],1-(3-aminopropyl)-2-pyrrolidine [7663-77-6],2-(aminomethyl)-ethy-1pyrrolidine [26116-12-1],(+)-S-2-(methoxymethyl)pyrrolidine [63126-47-6], 1-(pyrrolidineocarbonylmethyl) piperazine [339890-45-4].

[0185] C. Amino Groups Derived from Monomer 3

[0186] The reaction of Monomer 3 with a preformed diaminochloro(triazine) provides the final step in the synthesis of N², N⁴,N⁶-tris(amino-substituted)-1,3,5-triazines. Thus, for bonding the thirdpendant amino group to the triazine core, the Monomer 3 used andproposed consisted of amines, specifically alkyl (C₁-C₁₂, straight chainor branched), cycloalkyl (C₃-C₁₀ ring size), azacyclo (C₂-C₁₀), andbenzyl amine derivatives. The ring of these cycloalkyl-, azacycloamine,and phenyl ring of the benzyl derivatives can be optionally substitutedwith one or more moieties, or a combination of moieties such as groupsas alkyl, alkenyl, alkynyl, phenyl, benzyl, halo, cyano, nitro, hydroxy,thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio, arylthio, amino, alkylamino, aryl amino, acyl, carboxyl, amido, sulfonamido, sulfonyl,sulfate, sulfonic acid, morpholino, thiomorpholino, piperazinyl,pyridyl, thienyl, furanyl, pyrroyl, pyrazoyl, phosphate, phosphonicacid, phosphonate and the like.

[0187] In addition, any monomer described that has a stereocenterincludes its enantiomers, diastereomers, and optical isomers whether inenantiomeric or racemic forms, or mixtures of stereoisomers. This is toinclude all of the 1,3,5-triazine derivatives and their stereoisomerspresented herein that are formed as a result of using optically active,scalemic or racemic monomers.

[0188] Specific examples of Monomer 3 used to attach the third pendantamino group in the synthesis of N², N⁴,N⁶-tris(amino-substituted)-1,3,5-triazines, and their corresponding[Chemical Abstract Registry numbers] used in the synthesis of the N²,N⁴, N⁶-tris(amino-substituted)-1,3,5-triazine derivatives include, butare not limited to, ethylamine [75-04-07], cyclohexanemethylamine[3128-02-08], tert-butylamine [75-64-9], furfurylamine [617-89-0],benzylamine [100-46-9], 2,2,2-trifluroethylamine [753-90-2],cyclooctylamine [5452-37-9], N,N-dimethylethylenediamine,cyclohexylamine [108-91-8], cyclopentylamine [1003-03-8],1-(2-aminoethyl)-piperidine, [26116-12-1], 1-acetylpiperazine[13096-96-3], pyrrolidine [123-75-1], 1-piperonylpiperazine[32231-06-4]hexamethyleneimine [111-49-9], 1-(2-pyridyl)piperazine[34803-66-2], decahydroquinoline (cis/trans) [2051-28-7],1-methylpiperazine [109-01-3], 1-(3-aminopropyl)-imidazole [5036-48-6],ethyl 1-piperazine carboxylate [120-43-4], 4-methylcyclohexylamine(cis/trans) [6321-23-9], 1 -(3-aminopropyl)-2-pyrrolidine [7663-77-6],2-(aminomethyl)-ethyl-pyrrolidine [26116-12-1], (+)-S-2-(methoxymethyl)pyrrolidine [63126-47-6], 1-(pyrrolidineocarbonylmethyl) piperazine[339890-45-4].

[0189] In addition to the parallel synthetic procedures used to preparethe compounds of Table 2, Table 1 also provides other exemplary triazinecompounds of the present invention, which were synthesized individuallyrather than using parallel syntheses. The complete preparation andproperties of these compounds are presented in the Examples, wheredetails of the synthetic procedures used are provided. The syntheticprocedures for these compounds involve both the substitution of chloridegroups on cyanuric chloride, as well as various chemical modificationsof these groups once bonded to the trizine core. In particular, thisinvention also encompasses salts of the neutral triazine compounds, asprovided in the Examples and the Tables.

[0190] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0191] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0192] wherein:

[0193] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0194] X¹ is selected from m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂, m-SO₂R¹,or m-SO₂OR¹, or X¹ and X² together is a fused benzene, pyridine, ordioxane ring;

[0195] X² is selected from p-OR¹, p-SR¹, p-NR¹ ₂, p-OM, or p-SM, whereinM is selected from Li, Na, K, Mg, or Ca;

[0196] Y¹ is selected from cycloalkyl with up to 10 carbon atoms; linearor branched alkyl with up to 10 carbon atoms; CH₂R², wherein R² is acycloalkyl with up to 10 carbon atoms; or

[0197] wherein n is 1 or 2;

[0198] AY² is selected from a halogen or OR¹, or

[0199] A is NR¹ and Y² is selected from R¹,

[0200] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0201] In a further aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0202] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0203] wherein:

[0204] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; cycloalkyl with up to 10carbon atoms; or aryl;

[0205] E is CH or N;

[0206] n is an integer from 0 to 3;

[0207] X¹ is selected from —H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,m-SO₂R¹, or m-SO₂OR¹, or X¹ and X² together is a fused benzene orpyridine ring;

[0208] X² is selected from —H, o-Cl, o-Br, p-OR¹, p-SR¹, p-NR¹ ₂, p-F,p-Cl, p-Br, p-CF₃, p-C(O)OR¹, p-OM, or p-SM, wherein M is selected fromLi, Na, K, Mg, or Ca;

[0209] A is selected from NR¹ or O, wherein Y¹ is selected fromcycloalkyl with up to 10 carbon atoms, linear or branched alkyl with upto 10 carbon atoms, or

[0210] when A is NR¹, and wherein Y¹ is selected from R¹ or CH₂R¹ when Ais O, or AY¹ is selected from a halogen,

[0211] and

[0212] DY² is a halogen, or D is NR¹ and Y² is selected from

[0213] or (CHR¹)_(x)NR¹ ₂, wherein x is an integer from 1 to 6.

[0214] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0215] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0216] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0217] wherein:

[0218] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; cycloalkyl with up to 10carbon atoms; aryl; or (CH₂)_(x)CN, wherein x is an integer from 0 to 6;

[0219] E is CH or N;

[0220] n is an integer from 0 to 3;

[0221] X¹ is selected from —H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,m-SO₂R¹, m-SO₂OR¹, m-NC(O)R¹, or o-F, or X¹ and X² together is a fusedbenzene, pyridine, or dioxane ring;

[0222] X² is selected from —H, o-Cl, o-Br, o-CF₃, o-R¹, p-OR, p-SR¹,p-NR¹ ₂, p-F, p-Cl, p-Br, p-CF₃, p-CN, p-C(O)OR¹, p-NC(O)R¹,p-(4-morpholinyl), or p-(4-methyl-1-piperazinyl);

[0223] AY¹ is a halogen, or A is NR¹ or O and Y¹ is selected fromcycloalkyl with up to 10 carbon atoms, cycloalkyl with up to 10 carbonatoms substituted with R¹, linear or branched alkyl with up to 10 carbonatoms, CH₂R¹, (CHR¹)_(y)OR¹, wherein y is an integer from 1 to 6,

[0224] or AY¹ together are

[0225] wherein x is an integer from 3 to 5; and

[0226] DY² is a halogen, or D is NR¹ and Y² is selected from

[0227] cycloalkyl with up to 10 carbon atoms, cycloalkyl with up to 10carbon atoms substituted with R¹, linear or branched alkyl with up to 10carbon atoms, CH₂R¹,

[0228] wherein x is an integer from 3 to 5,

[0229] CH₂CF₃, (CHR¹)_(z)Z¹, wherein z is an integer from 1 to 6, and Z¹is selected from NR¹ ₂,

[0230] wherein x is an integer from 3 to 5,

[0231] or NY²R¹ together is selected from

[0232] wherein Z² is selected from R¹, C(O)R¹, C(O)OR¹, pyridinyl, aryl,

[0233] wherein q is an integer from 0 to 6.

[0234] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0235] As an additional aspect of this invention includes compounds ofthe present invention include, but are not limited to, those having thefollowing formula:

[0236] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0237] wherein:

[0238] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0239] X¹ is selected from H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,m-SO₂R¹, or m-SO₂OR¹;

[0240] X² is selected from o-R¹, p-OR¹, p-SR¹, p-NR¹ ₂, p-OM, or p-SM,wherein M is selected from Li, Na, K, Mg, or Ca;

[0241] Y¹ is selected from cycloalkyl with up to 10 carbon atoms or

[0242] and

[0243] Y²is selected from linear or branched alkyl with up to 10 carbonatoms, cycloalkyl with up to 10 carbon atoms, or

[0244] and R² is —H; or NY²R² together is selected from

[0245] wherein x is an integer from 3 to 5,

[0246] wherein q is an integer from 0 to 6, or

[0247] wherein Z² is selected from R¹ or

[0248] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0249] In still another aspect of this invention, compounds of thepresent invention include, but are not limited to, those having thefollowing structural formula:

[0250] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0251] wherein:

[0252] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0253] X¹ is in each occurrence independently selected from —H, m-F,m-Cl, m-Br, m-I, m-CN, m-NO₂, m-SO₂R¹, or m-SO₂OR¹;

[0254] X² is in each occurrence independently selected from o-CH₃,p-OR¹,p-SR¹,p-NR¹ ₂, or p-OM or p-SM, wherein M is selected from Li, Na,K, Mg, or Ca;

[0255] Y¹ is selected from cycloalkyl with up to 10 carbon atoms;

[0256] wherein n is 1 or 2; or

[0257] and

[0258] Y² is selected from

[0259] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0260] These compounds and compositions presented above are not intendedto be limiting, but merely representative of the chemical structures andformulas encompassed by the present invention.

[0261] Pharmaceutically Acceptable Salts

[0262] For the proposed N², N⁴,N⁶-tris(amino-substituted)-1,3,5-triazines, the terms “non-toxic,pharmaceutically acceptable salt” or “pharmaceutically acceptable salt”refer to a salt or complex of the 1,3,5-triazine compounds that retainor enhance the biologically activity of the compounds described in thisinvention. Examples of salts are those that are derived from theinteraction of the 1,3,5-triazine compounds or derivatives and aninorganic (mineral acids) or organic acid, as well as compounds derivedfrom deprotonating an amine nitrogen of the triamine derivatives.

[0263] Examples of inorganic acids include, but are not limited to,hydrochloric acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid,nitric acid, nitrous acid, perchloric acid, chloric acid, hypochlorousacid, chlorous acid, phosphoric acid, sulfuric acid, sulfurous acid, andcarbonic acid. Examples of organic acids include, but are not limited toacetic acid, benzene sulfonic acid, benzoic acid, butanoic acid,camphorsulfonic acid, citric acid, ethane sulfonic acid, fumaric acid,glutaric acid, 2-hydroxy acetic acid acids (derivatives where alkylgroup is c=3-7 and hydroxy group is located accordingly), 2-hydroxyalkyl sulfonic acids (derivatives where alkyl group is c=3-7 and hydroxygroup is located accordingly), lactic acid, maleic acid, malic acidmalonic, methane sulfonic acid, naphthalene sulfonic acid, oxalic acid,palmitic acid, propanoic acid, phthalic acid, pyruvic acid, salicylicacid, stearic acid, succinic acid, tartaric acid, p-toluene sulfonicacid, and amino acids (e.g., alanine, N-acetylglycine, arginine,aspartic acid, glutamic acid, glycine, lysine, and phenylalanine).

[0264] Examples of salts described here include compounds that derivefrom a deprotonation reaction of an amine nitrogen of the triaminederivatives with a strong base, to form an amido salt, compound orcomplex. For example, these compounds include those that are derivedfrom the interaction or chemical reaction of the 1,3,5-triazinecompounds or derivatives acting as a Bronsted or Lewis acid and aninorganic or organic base to form an ionic and/or complexed speciesExamples of inorganic bases, include but not limited to, metallic basesor organometallic bases such as alkyllithiums or metal hydrides, wherethere is a metallic counterion include, but are not limited to,aluminum, barium, calcium, lithium, magnesium, potassium sodium, andzinc.

[0265] Examples of organic bases include, but are not limited to, alkyand aryl amines as well as ammonia. Included in this description aresalts formed from the combination or interaction/reaction of inorganicacids (e.g., Lewis acids) and metallic counterions and the1,3,4-triazine compounds or derivatives acting as a Bronsted or Lewisbase resulting in the formation of ionic and/or complexed species Forall salts and complexes as described above, these are to includehydrated or solvated forms of the compounds.

[0266] Additionally, this invention also encompasses salts of thesetriazine derivatives that are non-toxic and pharmaceutically acceptable,such as quaternary ammonium salts, for example [—N⁺R₂R′]X⁻, where the Rand R′ groups represent hydrogen or an organic group (such as alkyl,alkenyl, alkynyl, aryl, and the like) and the X group is a counter ion(halogen, hydroxide, alkoxide, thioalkoxide, or conjugate base of anorganic or inorganic acid). For all salts and complexes as describedabove, these are to include hydrated or solvated forms of the compounds.

[0267] In one aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0268] wherein

[0269] G is selected from NH or O;

[0270] Z is selected from H or

[0271] wherein X¹ is selected from F or Cl, and X² is selected fromOCH₃, NH₂, OC(O)CH₃, or OH;

[0272] A is selected from NR¹ or O;

[0273] Y¹ is selected from R¹,

[0274] B is selected from NR¹ or O; and

[0275] Y² is selected from

[0276]  wherein q is 0 or 1, E is selected from O or NR² wherein R² isselected from R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂;

[0277] or

[0278] wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and

[0279] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0280] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0281] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0282] wherein:

[0283] G is selected from NH or O;

[0284] X¹ is selected from F or Cl;

[0285] A is selected from NR¹ or O;

[0286] Y¹ is selected from R¹,

[0287] B is selected from NR¹ or O; and

[0288] Y² is selected from

[0289] wherein q is 0 or 1, E is selected from O or NR², and R² isselected from R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂;

[0290] wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and

[0291] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0292] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0293] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0294] wherein:

[0295] G is selected from NH or O;

[0296] X¹ is selected from F or Cl; and

[0297] E is selected from O or NR², wherein R² is selected from R¹, OR¹,C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂;

[0298] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0299] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0300] In still another aspect of this invention, compounds of thepresent invention include, but are not limited to, those having thefollowing formula:

[0301] wherein:

[0302] G is selected from NH or O;

[0303] X¹ is selected from F or Cl; and

[0304] B is selected from NR¹ or O; and

[0305] R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂;

[0306] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0307] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0308] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0309] wherein:

[0310] G is selected from NH or O;

[0311] Z is selected from H or

[0312] B is selected from NR¹ or O; and

[0313] R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂;

[0314] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0315] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0316] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0317] wherein:

[0318] Q is selected from NH₂,OC(O)R¹, or OH;

[0319] X¹ is selected from F or Cl;

[0320] B is selected from NR¹ or O; and

[0321] Y²is selected from

[0322] wherein q is 0 or 1, E is selected from O or NR², and R² isselected from R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂;

[0323] or

[0324] wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and

[0325] wherein R¹ is in each occurrence independently selected from H;or a linear or branched alkyl with up to 10 carbon atoms.

[0326] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0327] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0328] wherein

[0329] GZ is selected from NH₂,OH,

[0330] AY¹ is selected from OH, NH₂,

[0331] and

[0332] BY² is selected from OH,

[0333] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0334] In still another aspect of this invention, compounds of thepresent invention include, but are not limited to, those having thefollowing formula:

[0335] wherein

[0336] AY¹ is selected from OH, NH₂,

[0337] and

[0338] BY² is selected from OH,

[0339] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0340] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0341] wherein

[0342] AY¹ is selected from OH,

[0343] and

[0344] BY² is selected from OH,

[0345] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0346] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0347] wherein

[0348] AY¹ is selected from

[0349] and

[0350] BY2 is selected from

[0351] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0352] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0353] wherein

[0354] AY¹ is selected from

[0355] and

[0356] BY² is selected from

[0357] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0358] In still another aspect of this invention, compounds of thepresent invention include, but are not limited to, those having thefollowing formula:

[0359] wherein:

[0360] GZ is selected from NH₂, OH,

[0361] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0362] In another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0363] wherein:

[0364] GZ is selected from

[0365] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0366] In yet another aspect of this invention, compounds of the presentinvention include, but are not limited to, those having the followingformula:

[0367] wherein:

[0368] GZ is selected from

[0369] BY² is selected from

[0370] Compositions comprising compounds of this formula are alsoencompassed by the present invention, as well as mixtures orcombinations of compounds of this formula.

[0371] Table 1D presents further exemplary compounds of the presentinvention. The inclusion of compounds in this table is not to be seen aslimiting, rather compounds are provided in this table by way of example.

[0372] Additionally, compounds in accordance with the present inventioninclude the compounds listed in Table 1E. Again, these compounds, whichare discussed in the Examples provided herein, are to be considered asexemplary. TABLE 1E Representative Compounds According to the PresentInvention COMPOUND NUMBER COMPOUND NAME E1 4-Benzyloxy-3-chloro-phenylamine, E2 N-(4-Benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine, E3 N-(4-benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine, E4 4-[4-Cycloheptylamino-6-(methyl-piperidin-4-yl-amino)-[1,3,5]triazin-2-ylamino]-phenol, E5 4-{4-Cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-1,3,5]triazin-2-ylamino}-phenol, E6 2-Chloro-4-(4,6-dichloro-[1,3,5]triazin-2-ylamino)-phenol, E7 2-Chloro-4-(4-chloro-6-cycloheptylamino-[1,3,5]triazin-2-ylamino)-phenol,E8  N-(1-Benzyl-piperidin-4-yl)-N′-(3-fluoro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-diamine, E9 N-Cycloheptyl-N′-(4-methoxy-phenyl)-N″-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine, E106-Chloro-N-cyclopropyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine, E11N-Cyclopropyl-N′-(3-flouro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine, E12N-Cyclopropyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine, E136-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cyclopropyl-[1,3,5]triazine-2,4-diamine E14N-Cyclopropyl-N′-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine, E156-Chloro-N,N′-bis-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine, E16N-(1-Ethyl-pyrrolidin-2-ylmethyl)-N′,N″-bis-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine, E171-[4-(3-chloro-4-methoxyanilino)-cycloheptyl amino-1,3,5-triazin-2-yl]-2-azoloamymethanol, E18N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(4-methylpiperzino)-1,3,5-triazine-4,2-diamine, E193-[4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazin-2-yloxy]-2-ethyl-4H-4-pyranone, E201-[3-{4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazine-2-yloxy}piperidino]-1-ethanone, E21N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-isopropoxy-1,3,5-triazine-2,4-diamine, E22N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine, E231-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-3-ol], E241-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-4-ol], E25N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine, E26N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-4-piperidyloxy)-1,3,5-triazine-2,4-diamine, E27N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1,4-thiazinan-4-yl)-1,3,5-triazine-4,2-diamine, E28N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-fluorophenoxy)-1,3,5-triazine-4,2-diamine, E29N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-[2-(2- fluorophenoxy)ethoxy]-1,3,5-triazine-4,2-diamine, E30N²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(6-methyl- 2-pyrylmethoxy]-1,3,5-triazine-4,2-diamine, E31N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl- N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine, HCl salt, E326-methoxy-N-(3-Chloro-4-methoxy phenyl)-N′-cyclo heptyl- [1,3,5]triazine-2,4 diamine, E33 6-ethoxy-N-(3-Chloro-4-methoxyphenyl)-N′-cyclo heptyl- [1,3,5] triazine-2,4 diamine, E346-isopropoxy-N-(3-Chloro-4-methoxy phenyl)-N′-cyclo heptyl- [1,3,5]triazine-2,4 diamine.

III. Antiproliferative Activities

[0373] One embodiment of the present invention comprises methods andcompositions comprising the compounds of the present invention for thetreatment and prevention of conditions or diseases that have as anaspect of the disease or condition, unwanted cellular proliferationoccurring or are the result of cellular proliferation. For example, manyvascular diseases, such as cardiovascular diseases, organ transplantsequellae, vascular occlusive conditions including, but not limited to,neointimal hyperplasia, restenosis, transplant vasculopathy, cardiacallograft vasculopathy, atherosclerosis, and arteriosclerosis, arecaused by or have collateral damage due to unwanted cellularproliferation. Smooth muscle cell (SMC) hyperplasia is a major event inthe development of atherosclerosis and is also responsible for thesignificant number of failure rates following vascular procedures suchas angioplasty and coronary artery bypass surgery, particularly due torestenosis. Proliferation of arterial wall SMC in response to localinjury is a major feature of many vascular proliferative disorders.Neointimal hyperplasia is commonly seen after various forms of vascularinjury and a major component of the vein graft's response to harvest andsurgical implantation into high-pressure arterial circulation.Proliferation of SMC in response to local injury is a major feature ofvascular proliferative disorders such as atherosclerosis and restenosisafter angioplasty.

[0374] One aspect of the present invention relates to methods andcompositions for the treatment and prevention of smooth muscle cell(SMC) proliferation, preferably comprising compositions and compoundshaving cellular antiproliferative activity. These compounds andcompositions comprising such compounds are referred to asantiproliferative compounds or compositions. At least one activity ofone or more of these compounds is that the compound has the activity ofeffecting the synthesis of proteoglycans including induction andsynthesis of proteoglycans and active fragments of proteoglycans. Thus,one aspect of the activity of one or more of the compounds andcompositions of the present invention comprise molecules that induceHSPG production and that regulate SMC (smooth muscle cell)proliferation.

[0375] Compounds of the present invention that have at least theactivity of effecting cellular proliferation are shown in TABLE 3. Thecompounds shown in this Table have the activity of effecting cellularproliferation as measured by the assays taught herein. The inclusion ofcompounds in the categories of the Tables disclosed herein are not to beseen as limiting, in that compounds included in such Tables have atleast the activity shown for inclusion in the Table and may have more orother activities. Nor are the Tables to be seen as limiting in thatthese are the only compounds disclosed herein that have that activity,representative compounds are shown in the Tables that have at least thatparticular activity for inclusion in the Table. One or more compoundsdisclosed herein have at least an activity that has utility in treatmentof disease states.

[0376] Examples of compounds that show at least this activity andutility are shown in the following structure:

[0377] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0378] wherein:

[0379] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0380] X¹ is selected from m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂, M-SO₂R¹,or m-SO₂OR¹, or X¹ and X² together is a fused benzene, pyridine, ordioxane ring;

[0381] X² is selected from p-OR¹, p-SR¹, p-NR¹ ₂,p-OM, or p-SM, whereinM is selected from Li, Na, K, Mg, or Ca;

[0382] Y¹ is selected from cycloalkyl with up to 10 carbon atoms; linearor branched alkyl with up to 10 carbon atoms; CH₂R², wherein R² is acycloalkyl with up to 10 carbon atoms; or

[0383] wherein n is 1 or 2;

[0384] AY² is selected from a halogen or OR¹, or

[0385] A is NR¹ and Y² is selected from R¹,

[0386] Further examples of compounds that show at least this activityand utility are presented in Table 3, where compound activity is alsopresented. The activity scale used in Table 3 as follows (numbers areinclusive): “+++” represents IC₅₀ of less than about 3 μM; “++”represents IC₅₀ of between about 3 and about 7 μM; and “+” representsIC₅₀ of greater than about 7 μM. In addition, compounds, to includecompositions thereof, encompassed within the scope of structuresId-XIVd, respectively, or listed in Tables 1D and 1E may be likewiseemployed in this embodiment and/or aspect of the present invention.Further, any hydrogen atoms that are required for any atom to attain itsusual valence in a structure presented in Table 3, whether a carbon atomor a heteroatom, should be inferred if it is not specifically indicated.

[0387] In addition to the above compounds, the following compounds andcompositions comprising these compounds are active in ananti-proliferation assay (Perlecan). These compounds and compositionscomprising these compounds are, among other things, generally useful fortreating cardiovascular disorders associated with proliferativeactivity. Specifically, these compounds includeN²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine,andN²-cycloheptyl-N⁴-methyl-N⁴-(1-methyl-piperidin-4-yl)-N⁶-naphthalen-2-yl-1,3,5-triazine-2,4,6-triamine.Using the same activity scale used in Table 3, and discussed above, thefirst compound,N²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine, is characterized as a compound exhibitingmedium or moderate activity, while the second compound,N²-cycloheptyl-N⁴-methyl-N⁴-(1-methyl-piperidin-4-yl)-N⁶-naphthalen-2-yl-1,3,5-triazine-2,4,6-triamine,is characterized as a compound exhibiting high activity.

[0388] As used herein, when a proteoglycan is referred to, the entiremolecule or fragments are included therein. For example, perlecan refersto the entire perlecan molecule or fragments thereof. Differentfragments of perlecan may have the same or different effects on cellsand the effects may be the same as or different from the effects thatthe entire perlecan molecule has on cells. These fragments andactivities are contemplated in the present invention and compoundsincluded in the present invention may have at least one activity thatmodulates or effects the fragements' activities or the entire molecule'sactivities. Although the discussion herein refers specifically toperlecans it is important to note that the compositions, methods, andassays described herein are equally applicable in the context of otherproteoglycans, including HSPGs, and including but not limited to,chondroitan sulfates (e.g., A,B, and C), dermatan sulfates, syndecansand glypicans.

[0389] Methods for identifying the activity and screening for one ormore of these compounds or molecules that induce synthesis ofproteoglycans such as HSPG (heparan sulfate proteoglycan) are taught inU.S. patent application Ser. No. 10/091,357, which is incorporatedherein in its entirety. Assays of effects of compounds in vivo are alsotaught in the incorporated references and are known to those skilled inthe art. In general, methods comprise the addition of such compounds toassays and measurement of HSPG synthesis including, but not limited to,the production of syndecans, glypicans and perlecans, for example,syndecans 1, 2 and 4; and glypican-1. Other assays that can be used todetermine the activity of the compounds of the present invention includeother methods for measuring the induction of perlecan synthesis. Forexample, in one assay, perlecan is induced in cells by certain inducers,and the response is measured. Compounds of the present invention arethen added to a replicate assay and the effect on perlecan induction isdetermined. Using such methods, compounds are determined that can eitherinhibit perlecan, elevate induction of perlecan, or that have no effectat all. Those compounds that are effective as therapeutic agents canthen be used in animals, humans or patients with cellular proliferationdisease aspects, such as vascular-associated diseases or SMCproliferation pathologies.

[0390] Another assay for determining compounds having SMC effectscomprises adding a composition suspected of effecting SMC proliferationto smooth muscle cells in growth medium or serum-free medium. The changein cell proliferation can be measured by methods known to those skilledin the art, such as incorporation of labeled nucleotides into dividingcells' DNA, and compared to the proliferation of cells which are nottreated with the compound. Other measurements include directlydetermining levels of HSPG synthesis by measuring the amount or changein amount of HSPG such as with ELISA for HSPGs, and compared to theamount of HSPG synthesis in untreated cells. Other indirect or directmeasurement are contemplated by the present invention and are known tothose skilled in the art. For example, such methods include, but are notlimited to, measurement of RNA levels, RT-PCR, Northern blotting,Western blotting promoter-based assays to identify compounds that affectone or more proteoglycans and assays for proteoglycan biologicalactivity shown by recombinant proteins, partially purified proteins, orlysates from cells expressing proteoglycans in the presence or absenceof compounds of interest.

[0391] An assay for identifying and determining an activity of one ormore of the compounds of the present invention comprises identifyingcompounds that interact with the promoter regions of a gene, or interactand effect proteins that interact with the promoter region, and areimportant in the transcriptional regulation of the protein's expression.For example, if perlecan were the protein, in general, the methodcomprises a vector comprising regulatory sequences of the perlecan geneand an indicator region controlled by the regulatory sequences, such asan enzyme, in a promoter-reporter construct. The protein product of theindicator region is referred to herein as a reporter enzyme or reporterprotein. The regulatory region of the sequence of perlecan comprises arange of nucleotides from approximately −4000 to +2000 wherein thetranscription initiation site is +1, more preferably, from −2500 to+1200, most preferably, from −1500 to +800 relative to the transcriptioninitiation site.

[0392] Cells are transfected with a vector comprising thepromoter-reporter construct and then treated with one or morecompositions comprising at least one compound of the present invention.For example, the transfected cells are treated with a compositioncomprising a compound suspected of effecting the transcription ofperlecan and the level of activity of the perlecan regulatory sequencesare compared to the level of activity in cells that were not treatedwith the compound. The level of activity of the perlecan regulatorysequences are determined by measuring the amount of the reporter proteinor determining the activity of the reporter enzyme controlled by theregulatory sequences. An increase in the amount of the reporter proteinor the reporter enzyme activity shows a stimulatory effect on perlecan,by positively effecting the promoter, whereas a decrease in the amountor the reporter protein or the reporter enzyme activity shows a negativeeffect on the promoter and thus, on perlecan.

[0393] Additionally, the present invention comprises methods andcompositions that can be used with gene therapy methods and composition,such as those gene therapy methods comprising administering compositionscomprising nucleic acids that effect the synthesis or expression ofHSPGs, particularly perlecan. Such methods and compositions are taughtin U.S. patent application Ser. No. 10/091,357, incorporated herein byreference.

[0394] The present invention comprises methods and compositions formediating proteoglycan synthesis, expression and for the maintenance ofSMC in a quiescent state. Methods and compositions of the presentinvention comprise treatment and prevention of vascular diseases andpathologies related to celluar proliferation, such as SMC proliferation.Such methods and compositions comprise methods for inhibition of smoothmuscle cell (SMC) growth and proliferation, and for induction ofquiescence in smooth muscle cells. Embodiments of the present inventioncomprise methods and compositions for inducing proteoglycan synthesis,particularly HSPG synthesis and expression including, but not limitedto, the induction of HSPGs such as syndecans, glypicans and perlecans,and preferably perlecan synthesis and gene expression. Perlecan is amajor extracellular HSPG in the blood vessel matrix. It interacts withextracellular matrix proteins, growth factors and receptors. Perlecan isalso present in basement membranes other than blood vessels and in otherextracellular matrix structures.

[0395] The activities of the compounds included in the present inventioneffect cells or tissues to increase the synthesis of proteoglycans bythose cells or tissues or may act directly upon one or moreproteoglycans to modulate the biological activity or to increase thebiological stability of the proteoglycan itself, for example, of theprotein perlecan. Activities also included herein are ones that increasethe biosynthesis of one or more proteoglycans by increasing thetranscription of the poteoglycan gene, increasing the biologicalstability of the proteoglycan mRNA or increasing the translation ofproteoglycan mRNA into protein. Further activites include activities ofcompounds that can block or decrease the effects of agents or proteinsthat inhibit the activity of proteoglycans.

[0396] The present invention comprises methods and compositions for thetreatment and prevention of smooth muscle cell proliferation, includingvascular occlusive pathologies. Such methods comprise administration ofcompositions comprising compounds capable of inhibiting SMCproliferation, such as compositions comprising compounds disclosedherein that inhibit SMC proliferation. Administration of such compoundsthat are effective in inhibiting SMC proliferation are administered tohumans and animals suspected of having or who have, for example,vasculopathy or who have undergone angioplasty or other proceduresdamaging to the endothelium. Effective amounts are administered to suchhumans and animals in dosages that are safe and effective, including,but not limited to, the ranges taught herein. Routes of administrationinclude, but are not limited to, those disclosed herein. As disclosedherein, compositions comprising such compounds may be used inconjunction with other therapeutic agents or in methods comprising stepssuch as altered patient activities, including, but not limited to,changes in exercise or diet.

[0397] The compounds of the present invention are useful in thetreatment or prophylaxis of at least one cardiovascular disease in acell, tissue, organ, animal, or patient including, but not limited to,vascular occlusive lesions including atherosclerosis, transplantvasculopathy, cardiac allograft vasculopathy, restenosis, graftatherosclerosis after coronary transplantation, cardiac stun syndrome,myocardial infarction, congestive heart failure, stroke, ischemicstroke, hemorrhage, arteriosclerosis, atherosclerosis, restenosis,diabetic ateriosclerotic disease, hypertension, arterial hypertension,renovascular hypertension, syncope, shock, syphilis of thecardiovascular system, heart failure, cor pulmonale, primary pulmonaryhypertension, cardiac arrhythmias, atrial ectopic beats, atrial flutter,atrial fibrillation (sustained or paroxysmal), post perfusion syndrome,cardiopulmonary bypass inflammation response, chaotic or multifocalatrial tachycardia, regular narrow QRS tachycardia, specific arrythmias,ventricular fibrillation, His bundle arrythmias, atrioventricular block,bundle branch block, myocardial ischemic disorders, coronary arterydisease, angina pectoris, myocardial infarction, cardiomyopathy, dilatedcongestive cardiomyopathy, restrictive cardiomyopathy, valvular heartdiseases, endocarditis, pericardial disease, cardiac tumors, aordic andperipheral aneuryisms, aortic dissection, inflammation of the aorta,occulsion of the abdominal aorta and its branches, peripheral vasculardisorders, occulsive arterial disorders, peripheral atherloscleroticdisease, thromboangitis obliterans, functional peripheral arterialdisorders, Raynaud's phenomenon and disease, acrocyanosis,erythromelalgia, venous diseases, venous thrombosis, varicose veins,arteriovenous fistula, lymphederma, lipedema, unstable angina,reperfusion injury, post pump syndrome, ischemia-reperfusion injury, andthe like. Such methods can optionally comprise administering aneffective amount of a composition or pharmaceutical compositioncomprising at least one compound to a cell, tissue, organ, animal orpatient in need of such modulation, treatment or therapy.

[0398] Proteoglycan-associated diseases that are treatable with thecompounds of the present invention include, but are not limited to,hereditary multiple exostosis, mucopolysaccharidosis types I-III andVII, commonly known as Hurler's Syndrome, Hunter's Syndrome,Sanfilippo's Syndrome and Sly's Syndrome respectively, Alzheimer'sdisease, Simpson-Golabi-Behmel syndrome, fibroblast growth factorrelated disorders, herpes simplex virus, dengue fever, Parkinson'sdisease, renal disease, muscular dystrophy, Schwarts-Jampel syndrome,proteinuric glomerulopathies, myotonia and skeletal dysplasia,kyphoscoliosis, dyssegmental dysplasia, Silverman-Handmaker type,chondrodysplasia, periodontitis, rheumatoid and osteoarthritis,Gerstmann-Straussler syndrome, Creutzfeldt-Jakob disease, scrapie,carcinomas, Happle syndrome, macular dystrophy, bone diseases, cornealdiseases, leukocyte-mediated disease, collagen fibril assembly disorderand coronary heart disease and other vascular disorders.

IV. Glycosidase Modulation Activity

[0399] The present invention also comprises methods and compositionscomprising compounds described herein that have an activity associatedwith modulation of glycosidase enzymes and thus, effecting thesubstrates for such enzymes. Glycosidase enzymes and their activity withtheir substrates, such as proteoglycans or glycated proteins, areaspects of a variety of diseases such as vascular conditions, includingthose conditions discussed supra, proteoglycan-associated diseases,supra, associated diseases with vascular components, including but notlimited to, kidney disease, ischemic heart disease, cardiovasculardisease, generalized vascular disease, proliferative retinopathy, andmacroangeopathy, inflammatory diseases and metastatic diseases such ascancer, cellular proliferative conditions, and solid and blood bornetumors or other oncological conditions. Compounds described herein thathave an activity that effects the concentrations of substrates ofglycosidase enzymes are used in methods of treatment of such vascular,inflammatory, metastatic and systemic diseases.

[0400] An aspect of the present invention comprises methods andcompositions for the modulation of enzymes, such as glycosaminoglycandegrading enzymes, which effect or are effected by proteoglycan levels,amount or activity. For example, the present invention comprises methodsand compositions comprising compounds that modulate enzymes includingbut not limited to, heparanase, chondroitanase, heparan sulfateendoglycosidase, heparan sulfate exoglycosidase, polysaccharide lyases,keratinase, hyauronidase, glucanase, amylase, glycosidases, or otherproteoglycan degrading enzymes are useful for the treatment ofconditions such as diabetic vasculopathy, cancer, inflammatory diseases,autoimmune diseases and cardiovascular diseases. For example, thepresent invention comprises methods and compositions of compounds thatinhibit, impair or down-regulate the activity of proteoglycan degradingenzymes.

[0401] Proteoglycans such as HSPG are important components of thesubendothelial extracellular matrix and the basement membrane of bloodvessels. Rosenberg et al., 99 J. CLIN. INVEST. 2062-70 (1997). Basementmembranes are continuous sheets of extracellular matrix composed ofcollagenous and noncollagenous proteins and proteoglycans that separateparenchymal cells from underlying interstitial connective tissue. Theyhave characteristic permeabilities and play a role in maintaining tissuearchitecture.

[0402] In addition to HSPGs, the basal lamina consists predominantly ofa complex network of adhesion proteins, fibronectin, laminin, collagenand vitronectin. Wight et al., 6 CURR. OPIN. LIPIDOL. 326-334 (1995).Heparan sulfate (HS) is an important structural component of the basallamina. Each of the adhesion proteins interacts with HS side chains ofHSPGs within the matrix. Thus, HSPGs function as a barrier to theextravasation of metastatic and inflammatory cells. Cleavage of HS bythe endoglycosidase heparanase produced by metastatic tumor cells andinflammatory cells destroys the filtering properties of the lamina. Inaddition, the degradation of the HS may assist in the disassembly of theextracellular matrix and thereby facilitate cell migration by allowingblood borne cells to escape into the bloodstream. Vlodavsky et al., 12INVASION METASTASIS 112-127 (1992).

[0403] Heparanase activity has been described in a number of tissues andcell types including liver, placenta, platelets, fibroblasts,neutrophils, activated T and B-lymphocytes, monocytes, and endothelialcells (7-16). Nakajima et al., (31) CANCER LETT. 277-283 (1986);Nakajima et al., 36 J. CELL. BIOCHEM. 157-167 (1988); Ricoveri et al.,46 CANCER RES. 3855-3861 (1986); Gallagher et al., 250 BIOCHEM. J.719-726 (1988); Dempsey et al., 10 GLYCOBIOLOGY 467 (2000); Goshen etal., 2 MOL. HUM. REPROD. 679 (1996); Parish et al., 76 IMMUNOL CELLBIOL. 104-113 (1998); Gilat et al., 181 J. EXP. MED. 1929-1934 (1995);Graham, et al., 39 BIOCHEM. MOL. BIOL. INT. 56371 (1996); Pillarisettiet al., 270 J.BIOL.CHEM. 29760-29765 (1995). An important process intissue invasion by blood-borne tumor cells and white cells involvestheir passage through the vascular endothelial cell layer and subsequentdegradation of the underlying basal lamina or basement membranes andextracellular matrix with a battery of secreted proteases andglycosidases. Nakajinia et al., 220 SCIENCE 611-613 (1983); Vlodavsky etal.,12 INVASION METASTASIS 112-127 (1992).

[0404] Heparanase activity was shown to correlate with the metastaticpotential of animal and human tumor cell lines. Nakajima et al., 31CANCER LETT. 277-283 (1986); Nakajima et al., 212 PROG CLIN BIOL RES.113- 122 (1986); Freeman et al., 325 BIOCHEM. J. 229-237 (1997);Vlodavsky et al., 5 NAT. MED. 793-802 (1999); Hulett et al., 5 NAT MED.803-809 (1999). It is also known to regulate growth factor activity.Many growth factors remain bound to heparan sulfate in storage form andare disassociated by heparanase during angiogenesis, improving thesurvival rate of cancer cells.

[0405] Serum heparanase levels in rats were higher by more than an orderof magnitude after injection of the rats with highly metastatic mammaryadenocarcinoma cells. In addition, heparanase activity in the sera ofrats bearing MTLn3 tumors correlated well with the extent of themetastases. Moreover, serum/urine heparanase activity in cancer patientswas shown to be 2-4 fold increased in particular where tissue metastaseswere present. Because the cleavage of HS appears to be essential for thepassage of metastatic tumor cells and leukocytes through basementmembranes, studies of heparanase inhibitors provides the potential ofdeveloping a novel and highly selective class of anti-metastatic andanti-inflammatory drugs.

[0406] The present invention comprises methods and compositionscomprising compounds that modulate heparanase activity or the activityof other glycosidases, including, but not limited to enzymes withglycosaminoglycan activity such as chondroitinase, heparan sulfateendoglycosidase, heparan sulfate exoglycosidase, polysaccharide lyases,keratinase, hyaluranidase, glucanase, and amylase. Compounds of thepresent invention that have at least the activity of modulatingglycosidase enzyme activity are shown in TABLE 6. The compounds shown inthis Table have the activity of modulating glycosidase enzyme activityas measured by the assays taught herein. The inclusion of compounds inthe categories of the Tables disclosed herein are not to be seen aslimiting, in that compounds included in such Tables have at least theactivity shown for inclusion in the Table and may have more or otheractivities. Nor are the Tables to be seen as limiting in that these arethe only compounds disclosed herein that have that activity,representative compounds are shown in the Tables that have at least thatparticular activity for inclusion in the Table. One or more compoundsdisclosed herein have at least an activity that has utility in treatmentof disease states.

[0407] Examples of compounds that show at least this activity andutility are shown in the following formula:

[0408] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0409] wherein:

[0410] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0411] X¹ is selected from H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,M-SO₂R¹, or M-SO₂OR¹;

[0412] X² is selected from o-R¹, p-OR¹, p-SR¹, p-NR¹ ₂, p-OM, orp-SM,wherein M is selected from Li, Na, K, Mg, or Ca;

[0413] Y¹ is selected from cycloalkyl with up to 10 carbon atoms or

[0414] and

[0415] Y² is selected from linear or branched alkyl with up to 10 carbonatoms, cycloalkyl with up to 10 carbon atoms, or

[0416] and R² is —H; or NY²R² together is selected from

[0417] wherein x is an integer from 3 to 5,

[0418] wherein q is an integer from 0 to 6, or

[0419] wherein Z² is selected from R¹ or

[0420] Further examples of compounds that show at least this activityand utility are presented in Table 6, where compound activity is alsoshown. The activity scale used in Table 6 is as follows (numbers areinclusive): “+++” represents between about 70 and about 100% inhibition;“++” represents between about 30 and about 40% inhibition; and “+”indicates between 0 and about 30% inhibition, all at 5 μM compoundconcentration. In addition, compounds, to include compositions thereof,encompassed within the scope of structures Id-XIVd, respectively, orlisted in Tables 1D and 1E may be likewise employed in this embodimentand/or aspect of the present invention. Also note that any hydrogenatoms that are required for any atom to attain its usual valence in astructure presented in Table 6, whether a carbon atom or a heteroatom,should be inferred if it is not specifically indicated.

[0421] Compounds or compositions comprising such compounds that areeffective in modulating glycosidase enzyme activity are useful intreating and/or preventing cancer including, but not limited to,malignant and non-malignant cell growth, leukemia, acute leukemia, acutelymphoblastic leukemia (ALL), B-cell, T-cell or FAB ALL, acute myeloidleukemia (AML), chromic myelocytic leukemia (CML), chronic lymphocyticleukemia (CLL), hairy cell leukemia, myelodyplastic syndrome (MDS), alymphoma, Hodgkin's disease, a malignamt lymphoma, non-hodgkin'slymphoma, Burkitt's lymphoma, multiple myeloma, Kaposi's sarcoma,colorectal carcinoma, pancreatic carcinoma, nasopharyngeal carcinoma,malignant histiocytosis, paraneoplastic syndrome/hypercalcemia ofmalignancy, solid tumors, adenocarcinomas, sarcomas, malignant melanoma,hemangioma, metastatic disease, cancer related bone resorption, cancerrelated bone pain, and the like.

[0422] In another aspect of the present invention, the compoundsdisclosed herein are useful in modulating heparanase activity or theactivity of other glycosidases as a means for treating and preventingautoimmune diseases. Generally autoimmune disease results when (1) theimmune system mistakenly identifies a cell surface molecule on normaltissue as a foreign molecule (2) the synthesis and secretion ofchemokines, cytokines and lymphokines is not shut down after theeradication of the disease or (3) the immune system overreacts to theapparent infection and destroys vast quantities of surrounding normaltissue.

[0423] To be effective in an immune response, the immune effector cellsmust bind to the luminal/apical surface of the blood vessel walls. Thisis accomplished through the interaction of adhesion molecules on theimmune effector cells with their locally upregulated cognate receptorson the endothelial cells lining the vasculature near the site ofinfection. After binding to the apical surface and before entering theinflamed tissue, the immune effector cells must breach the basementmembrane (BM) and extracellular matrix (ECM) that surround the basalportion of the blood vessels and give the vessels their shape andstrength. The BM and ECM consists of structural proteins embedded in afiber meshwork consisting mainly of complex carbohydrate containingstructures (glycosaminoglycans), of which the main constituent isheparin sulfate proteoglycan (HSPG). In order to breach this barrier theimmune effector cell must weaken or destroy it, which is accomplishedthrough the local secretion of proteases and heparanase(s).

[0424] Thus, the inhibition of heparanase or the activity of otherglycosidases using the compounds of the present invention finds utitlityin treating arthritis and other autoimmune diseases. More specifically,the compounds of the present invention are useful in the treatment orprophylaxis of at least one autoimmune-related disease in a cell,tissue, organ, animal, or patient including, but not limited to,rheumatoid arthritis, juvenile rheumatoid arthritis, systemic onsetjuvenile rheumatoid arthritis, psoriatic arthritis, ankylosingspondilitis, gastric ulcer, seronegative arthropathies, osteoarthritis,inflammatory bowel disease, ulcerative colitis, systemic lupuserythematosis, antiphospholipid syndrome, iridocyclitis/uveitis/opticneuritis, idiopathic pulmonary fibrosis, systemic vasculitis/wegener'sgranulomatosis, sarcoidosis, orchitis/vasectomy reversal procedures,allergic/atopic diseases, asthma, allergic rhinitis, eczema, allergiccontact dermatitis, allergic conjunctivitis, hypersensitivitypneumonitis, transplants, organ transplant rejection, graft-versus-hostdisease, systemic inflammatory response syndrome, sepsis syndrome, grampositive sepsis, gram negative sepsis, culture negative sepsis, fungalsepsis, neutropenic fever, urosepsis, meningococcemia,trauma/hemorrhage, bums, ionizing radiation exposure, acutepancreatitis, adult respiratory distress syndrome, rheumatoid arthritis,alcohol-induced hepatitis, chronic inflammatory pathologies, Crohn'spathology, sickle cell anemia, diabetes, nephrosis, atopic diseases,hypersensitity reactions, allergic rhinitis, hay fever, perennialrhinitis, conjunctivitis, endometriosis, asthma, urticaria, systemicanaphalaxis, dermatitis, pernicious anemia, hemolytic disesease,thrombocytopenia, graft rejection of any organ or tissue, kidneytranslplant rejection, heart transplant rejection, liver transplantrejection, pancreas transplant rejection, lung transplant rejection,bone marrow transplant (BMT) rejection, skin allograft rejection,cartilage transplant rejection, bone graft rejection, small boweltransplant rejection, fetal thymus implant rejection, parathyroidtransplant rejection, xenograft rejection of any organ or tissue,allograft rejection, anti-receptor hypersensitivity reactions, Gravesdisease, Raynoud's disease, type B insulin-resistant diabetes, asthma,myasthenia gravis, -meditated cytotoxicity, type III hypersensitivityreactions, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy,monoclonal gammopathy, and skin changes syndrome), polyneuropathy,organomegaly, endocrinopathy, monoclonal gammopathy, skin changessyndrome, anti-phospholipid syndrome, pemphigus, scleroderma, mixedconnective tissue disease, idiopathic Addison's disease, autoimmunehemolytic anemia, autoimmune hepatitis, idiopathic pulmonary fibrosis,scleroderma, diabetes mellitus, chronic active hepatitis, vitiligo,vasculitis, post-MI cardiotomy syndrome, type IV hypersensitivity,contact dermatitis, hypersensitivity pneumonitis, allograft rejection,granulomas due to intracellular organisms, drug sensitivity,metabolic/idiopathic, Wilson's disease, hemachromatosis,alpha-1-antitrypsin deficiency, diabetic retinopathy, hashimoto'sthyroiditis, osteoporosis, hypothalamic-pituitary-adrenal axisevaluation, primary biliary cirrhosis, thyroiditis, encephalomyelitis,cachexia, cystic fibrosis, neonatal chronic lung disease, chronicobstructive pulmonary disease (COPD), familial hematophagocyticlymphohistiocytosis, dermnatologic conditions, psoriasis, alopecia,nephrotic syndrome, nephritis, glomerular nephritis, acute renalfailure, hemodialysis, uremia, toxicity, preeclampsia, arilylosingspondylitis, Behcet's disease, bullous pemphigoid, cardiomyopathy,celiac sprue-dermnatitis, chronic fatigue immune dysfunction syndrome(CFIDS), chronic inflammatory demyelinating polyneuropathy,Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, coldagglutinin disease, discoid lupus, essential mixed cryoglobulinemia,fibromyalgia-fibromyositis, Graves' disease, Guillain-Barré, Hashimoto'sthyroiditis, idiopathic thrombocytopenia purpura (ITP), IgA nephropathy,insulin dependent diabetes, juvenile arthritis, lichen planus, ménière'sdisease, multiple sclerosis, pemphigus vulgaris, polyarteritis nodosa,Cogan's syndrome, polychondrfits, polyglandular syndromes, polymyalgiarheumatic a, polymyositis and dermatomyositis, primaryagammaglobulinemia, Raynaud's5 phenomenon, Reiter's syndrome, rheumaticfever, Sjögren's5 syndrome, stiff-man syndrome, Takayasu arteritis,temporal arteritis/giant cell arteritis, Wegener's granulomatosis; okt3therapy, anti-cd3 therapy, cytokine therapy, chemotherapy, radiationtherapy (e.g., including but not limited toasthenia, anemia, cachexia,and the like), chronic salicylate intoxication, and the like.

[0425] Compounds having heparanase activity inhibition, that areeffective for example, in treatment of cancer and autoimmune disease,can be determined using assays such as those disclosed in U.S. patentapplication Ser. No. 09/952,648, which is incorporated herein in itsentirety. Such assays, which are used for measurement of cellular andenzymatic activities, both qualitatively and quantitatively, and inmethods for diagnosing metastases, metastatic potential and inflammatorystates, are performed with and without the addition of at least one ofthe compounds of the present invention to determine the activity of thecompound. Existing heparanase assays are taught in Goshen et al., 2 MOL.HUM. REPROD. 679-84 (1996); Nakajima et al., 31 CANCER LETT. 277-83(1986); and Vlodasky et al., 12 INVASION METASTASIS 112-27 (1992);Freeman and Parish, 325 BIOCHEM. J. 229-37 (1997); Kahn and Newman, 196ANAL. BIOCHEM. 373-76 (1991).. Solid-phase heparanase assays have alsobeen developed where chemically and biosynthetically radiolabeledheparin and HS chains were attached to a solid support, with release ofradiolabel from the solid support being a measure of enzyme activity.Assays using such procedures are taught in U.S. Pat. No. 4,859,581,which is entirely expressly herein incorporated by reference.

[0426] In general, a preferred assay comprises attaching one of abinding partner to a substrate for the enzyme to be measured, formingthe substrate-binding partner. Incubation with a sample comprising theenzyme to be measured allows for activity by the enzyme to be measuredin a reaction mixture. A portion or the whole reaction mixture,depending on the amount needed, is then mixed with the complementarybinding partner, so that the binding partners are bound together. Thisis the first binding reaction. After incubating to allow for binding,washings are performed. A complementary binding partner, complementaryto the first binding partner attached to the substrate, is added. Thiscomplementary binding partner may or may not be the same as the firstcomplementary binding partner. This is the second binding reaction. Thecomplementary binding partner in the second binding reaction is labeledin a manner that is detectable. For example, the complementary bindingpartner is labeled with an enzyme that causes a detectable color changewhen the appropriate reaction conditions exist. The difference betweenthe activity of the enzyme in the presence of a compound and the absenceof compound is used to determine the activity of the compound.

[0427] An example of a heparanase assay comprises the following steps. Acomposition comprising biotin-HS (heparan sulfate) is mixed with abiological sample such as a tumor sample, bodily fluid, or other fluidsuspected of having heparanase activity, to form a reaction mixture.This sample may be pretreated to remove contaminating or reactivesubstances such as endogenous biotin. A control portion for thisreaction mixture does not contain a compound of the present invention,whereas a test portion contains one or more compounds disclosed herein.After incubation, an aliquot or portion of the reaction mixture portionsis removed and placed in a biotin-binding plate. The biotin-bindingplate comprises any means for binding biotin, preferably to a solidsurface. See WO 02/23197, which is entirely expressly incorporatedherein by reference. After washing with buffers, a streptavidin-enzymeconjugate is added to the biotin-binding plate. Reagents for the enzymeare added to form a detectable color product. For example, a decrease incolor formation, from a known standard, indicates there was heparanaseactivity in the sample. The difference between the activity of theenzyme in the presence of a compound and the absence of compound is usedto determine the activity of the compound.

[0428] Using the above assays or those taught in the Examples herein,the amount of enzyme activity in a sample can be determined and theactivities of compounds of the present invention can be determined. Forexample, a composition comprising a compound of the present invention isadded to a known amount of heparanase either before or during theincubation of the heparanase and its substrate-binding partner. If thecompound alters the activity of the heparanase, the assay methods of thepresent invention will show a change in the amount of detectable label.Such assays are used for high throughput determination of the activityof compounds. See WO 02/23197, which is entirely expressly incorporatedherein by reference.

[0429] The activities of the compounds included in the present inventionmodulate the activity of glycosidases, either positively or negatively,include effects on the glycosidases either directly or indirectly. Thecompounds may modulate the synthesis of glycosidases by cells or tissuesor may act directly upon one or more glycosidases to modulate thebiological activity or the biological stability of the enzyme itself,for example, heparanase. Activities also included herein are ones thatincrease the biosynthesis of one or more glycosidases by increasing thetranscription of the glycosidase gene, increasing the biologicalstability of the glycosidase mRNA or increasing the translation ofglycosidase mRNA into protein. Further activites include activities ofcompounds that can block or decrease the effects of agents or proteinsthat inhibit the activity of glycosidases. Additionally, acitivities areincluded that effect the substrates for the glycosidases, such as thosediscussed supra in relation to proteoglycans, or effect the bindingparameters of the enzyme with its substrate, cofactors or stimulatory orinhibitory factors.

[0430] The present invention comprises methods and compositions for thetreatment and prevention of diseases or conditions that present orresult from glycosidase activity. Such methods comprise administrationof compositions comprising compounds capable of modulating heparanaseactivity, such as compositions comprising compounds disclosed hereinthat inhibit heparanase activity. Administration of such compounds thatare effective in modulating heparanase activity are administered tohumans and animals suspected of having or who have, for example,inflammatory conditions, autoimmune disease or diabetic vasculopathy.Effective amounts are administered to such humans and animals in dosagesthat are safe and effective, including, but not limited to, the rangestaught herein. Routes of administration include, but are not limited to,those disclosed herein. As disclosed herein, compositions comprisingsuch compounds may be used in conjunction with other therapeutic agentsor in methods comprising steps such as altered patient activities.

V. Inflammation Modulation

[0431] An embodiment of the present invention comprises methods andcompositions comprising compounds of the present invention for thetreatment and prevention of conditions or diseases that have as anaspect of the disease or condition, inflammation. An aspect of thepresent invention is directed to methods and compositions comprisingcompounds that are effective in inhibiting inflammation, particularlyinflammation associated with the accumulation or presence of glycatedproteins or AGE. The activity of modulating inflammation includes, butis not limited to, inhibiting inflammation and/or its associated cellactivation by glycated proteins or AGE, blocking the glycation ofproteins, blocking AGE interactions with receptors, blocking AGE-inducedsignaling or signaling-associated inflammatory responses, cytokineinduction, synthesis or release, AGE formation or AGE cross-linking.

[0432] The present invention also provides compositions for and methodsof treatment of biological conditions including, but not limited to,vascular complications of type I and type II diabetic-inducedvasculopathies, other vasculopathies, microangiopathies, renalinsufficiency, Alzheimer's syndrome, and inflammation-induced diseasessuch as atherosclerosis. Other inflammatory related diseases include,but are not limited to, inflammatory diseases of the joint such asrheumatoid arthritis, osteoarthritis, autoimmune diseases such as thosetaught supra, streptococcal cell-wall induced arthritis,adjuvant-induced arthritis, bursitis; inflammatory diseases of thethyroid such as acute, subacute and chronic thyroiditis, pelvicinflammatory disease, hepatitis; inflammatory bowel diseases such asCrohn's disease and colitis; neuroinflammatory diseases such as multiplesclerosis, abscess, meningitis, encephalitis, and vasculitis;inflammatory diseases of the heart such as myocarditis, chronicobstructive pulmonary disease, atherosclerosis, pericarditis;inflammatory diseases of the skin such as acute inflammatory dermatoses(urticaria (hives), spongiotic dermatitis, erythema multiforme (emminor), Stevens-Johnson syndrome (sjs, em major), toxic epidermalnecrolysis (ten) and chronic inflammatory dermatoses (psoriasis, lichenplanus, discoid lupus erythematosus, acne vulgaris); inflammatorydiseases of the eye such as uveitis, allergic conjunctivitis, cornealinflammation, intraocular inflammation, iritis; laryngitis and asthma.

[0433] The compounds of the present invention have utility in inhibitinginflammation and/or its associated cell activation by glycated proteinsor AGE. Pharmacological inhibition of AGE-induced cell activationprovides the basis for therapeutic intervention in many diseases,notably in diabetic complications and Alzheimer's disease. Therapeuticapproaches for inhibition of AGE-induced inflammation include, but arenot limited to, blocking the glycation of proteins, blocking AGEinteractions with receptors and blocking AGE-induced signaling orsignaling-associated inflammatory responses.

[0434] At least one activity of some of the compounds of the presentinvention is to block AGE effects by inhibiting AGE-induced signaling.The sequence of these signaling events leading to inflammation are notclear, but inhibition of these signaling events leads to reduced or noinflammatory results. Compounds that block AGE-induced up-regulation ofinflammatory molecules were determined using screening assays. Otheraspects of the present invention comprise methods and compositionscomprising compounds that block glycated protein-induced inflammation.Some compounds may effect AGE formation or AGE cross-linking.

[0435] At least one activity of some of the compounds of the presentinvention is to block AGE effects by inhibiting reactions with receptorsof AGE and such activities are also contemplated by the methods of thepresent invention for treatment of related pathologies. For example,RAGE, a known receptor for AGE, is a therapeutic target. Blocking RAGEinhibited AGE-induced inflammation. Prior to use of the compounds of thepresent invention, the multiple functions of RAGE and possible long termside effects of accumulated AGE in plasma, have prevented this method oftreatment from being implemented. However, using the methods andcompositions of the present invention, more specific inhibitorycompounds can be used for treatments and overcome the current problemswith treatments that target receptors.

[0436] Compounds of the present invention that have at least theactivity of modulating inflammation activity are shown in TABLE 5. Thecompounds shown in this Table have the activity of modulatinginflammation activity as measured by the assays taught herein. Theinclusion of compounds in the categories of the Tables disclosed hereinare not to be seen as limiting, in that compounds included in suchTables have at least the activity shown for inclusion in the Table andmay have more or other activities. Nor are the Tables to be seen aslimiting in that these are the only compounds disclosed herein that havethat activity, representative compounds are shown in the Tables thathave at least that particular activity for inclusion in the Table. Oneor more compounds disclosed herein have at least an activity that hasutility in treatment of disease states.

[0437] Examples of compounds that show at least this activity andutility are shown in the following formula:

[0438] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0439] wherein:

[0440] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; cycloalkyl with up to 10carbon atoms; aryl; or (CH₂)_(x)CN, wherein x is an integer from 0 to 6;

[0441] E is CH or N;

[0442] n is an integer from 0 to 3;

[0443] X¹ is selected from —H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,m-SO₂R¹, M-SO₂OR¹, m-NC(O)R¹, or o-F, or X¹ and X² together is a fusedbenzene, pyridine, or dioxane ring;

[0444] X² is selected from —H, o-Cl, o-Br, o-CF₃, o-R¹,p-OR¹, p-SR p-NR¹₂,p-F, p-Cl, p-Br, p-CF₃, p-CN, p-C(O)OR¹, p-NC(O)R¹, p-(4-morpholinyl),or p-(4-methyl-1-piperizinyl);

[0445] AY¹ is a halogen, or A is NR¹ or O and Y¹ is selected fromcycloalkyl with up to 10 carbon atoms, cycloalkyl with up to 10 carbonatoms substituted with R¹, linear or branched alkyl with up to 10 carbonatoms, CH₂R¹, (CHR¹)_(y)OR¹, wherein y is an integer from 1 to 6,

[0446] or AY¹ together are

[0447] wherein x is an integer from 3 to 5; and

[0448] DY² is a halogen, or D is NR¹ and Y² is selected from

[0449] cycloalkyl with up to 10 carbon atoms, cycloalkyl with up to 10carbon atoms substituted with R¹, linear or branched alkyl with up to 10carbon atoms, CH₂R¹,

[0450] wherein x is an integer from 3 to 5,

[0451] CH₂CF₃, (CHR¹)_(z)Z¹, wherein z is an integer from 1 to 6, and Z¹is selected from NR¹ ₂,

[0452] wherein x is an integer from 3 to 5,

[0453] or NY²R¹ together is selected from

[0454] wherein Z² is selected from R¹, C(O)R¹, C(O)OR¹, pyridinyl, aryl,

[0455] wherein q is an integer from 0 to 6.

[0456] Further examples of compounds that show at least this activityand utility are presented in Table 5, where compound activity is alsoshown. The activity scale used in Table 5 is as follows (numbers areinclusive): “++++” represents between 0 and about 25% of IL6 productioncompared to cells that did not receive compound (or per cent of controlIL6 production); “+++” represents between about 25 and about 50% ofcontrol IL6 production; “++” represents between about 50 and about 75%of control IL6 production; and “+” represents between about 75 and 100%of control IL6 production. The note “n.d.” indicates that the activityof the compound was not determined in the given assay. Further note thatany hydrogen atoms that are required for any atom to attain its usualvalence in a structure presented in Table 5, whether a carbon atom or aheteroatom, should be inferred if it is not specifically indicated.

[0457] In addition to the above compounds, the compounds shown in Table7, and compositions comprising these compounds, also exhibit theactivity of modulating inflammation activity as measured by the assaystaught herein. The activity scale used in Table 7 is as follows (numbersare inclusive): “+++” represents between about 85 to 100% inhibition ofIL6 production in the presence of AGE or TNF, as compared to cells thatdid not receive any compound; “++” represents between about 65 and about85% inhibition of IL6 production in the presence of AGE or TNF; and “+”represents between about 50 and about 65% inhibition of IL6 productionin the presence of AGE or TNF. In addition, compounds, to includecompositions thereof, encompassed within the scope of structuresId-XIVd, respectively, or listed in Tables 1D and 1E may be likewiseemployed in this embodiment and/or aspect of the present invention. Asbefore, the inclusion of compounds in the categories of the Tablesdisclosed herein are not to be seen as limiting, in that compoundsincluded in such Tables have at least the activity shown for inclusionin the Tables and may have more or other activities. Nor are the Tablesto be seen as limiting in that these are the only compounds disclosedherein that have that activity, representative compounds are shown inthe Tables that have at least that particular activity for inclusion inthe Table. One or more compounds disclosed herein have at least anactivity that has utility in treatment of disease states.

[0458] Enhanced formation and accumulation of glycated proteins and AGEare thought to play a major role in the pathogenesis of diabeticcomplications, and atherosclerosis, leading to the development of arange of diabetic complications including nephropathy, retinopathy andneuropathy. There is ample in vivo evidence that suggests thatdiabetes-related complications can be reduced by 1) preventing glycationof proteins, 2) by breaking the cross-links in glycated proteins or 3)by blocking glycated protein interaction with receptors. Despite theimportance of AGE in the pathogenesis of diabetic microangiopathies,there are no currently available medications known to block AGEformation.

[0459] Endothelium is the target organ of damage in diabetes. See Laightet al., 15 DIABETES METAB. RES. REV. 274-82 (1999); Stehouwer et al., 34CARDIOVASC. 55-68 (1997). Up-regulation of molecules involved inendothelial inflammation, such as IL-6 and monocyte chemoattractantprotein-1 (MCP-1) leads to endothelial dysfunction and vasculopathy. SeeStehouwer et al., 34 CARDIOVASC. 55-68 (1997); Libby, 247 J. INTERN.MED. 349-58 (2000); Van Lente, 293 CLINICA. CHIMICA. ACTA. 31-52 (2000).

[0460] IL-6 is a pro-inflammatory cytokine that is known to play a keyrole in the pathogenesis of diabetes and atherosclerosis. See Horii etal., 39 KIDNEY INT. SUPPL. 71-5 (1993); Huber et al., 19ARTERIOSCLERTHROMB. VASC. BIOL. 2364-67 (1999); Shikano et al., 85NEPHRON 81-5 (2000); Pickup et al., 8(67) LIFE SCI. 291-300 (2000). IL-6also promotes the growth of renal mesangial cells thus contributing tonephropathy. See Kado et al., 36 ACTA. DIABETOL. 67-72 (1999). The serumIL-6 level in diabetic subjects was significantly higher than in normalhealthy controls (3.48±3.29 pg/mi vs 0.784±0.90 pg/ml, mean ±SD). Inaddition the urinary IL-6 level is a good indicator of diabeticnephropathy. Serum IL-6 is useful in the evaluation of atherosclerosisand nephropathy.

[0461] MCP-1, another pro-inflammatory cytokine is found highlyexpressed in human atherosclerotic lesions and postulated to play acentral in monocyte recruitment into the arterial wall and developinglesions. See Libby, 247 J. INTERN. MED. 349-58 (2000). Recent resultsshow that MCP-1 is also a key pathogenic molecule in diabeticnephropathy. See Eitner et al., 51 KIDNEY INT. 69-78 (1997); Banba etal. 58 KIDNEY INT. 684-90 (2000). Glycated albumin stimulatesendothelial production of IL-6 and MCP-1. The effects of glycatedalbumin on IL-6 production are comparable to that of TNFα, a knowninducer of IL-6. These cytokines are known to be factors in vasculardiseases.

[0462] The activity of the compounds of the present invention ininhibiting glycated protein- and AGE-induced inflammation can bedetermined using the assays described herein and in U.S. patentapplication Ser. No. 10/026,335, which is herein incorporated in itsentirety. Such assays comprise measurement of the specific activity ofbiological components involved in a known cellular response. The assaysprovide a measurable response in which the activity of the compounds isdetermined. One assay comprises measurement of the effects of compoundson an inflammatory response by cells to the presence of a stimulatingagent. Yet another assay comprises endothelial cells that are stimulatedby the addition of a glycated protein, the stimulating agent. Theendothelial cells respond by producing specific cytokines. The amount ofcytokines produced are determined by measurement protocols known tothose skilled in the art. The compounds of the present invention arethen added to the assay and the production of cytokines is measured.From the comparison of the assay without the compound with the assaywith the compound, the biological effect of the compound can bedetermined. The compound may have an inhibitory effect, a stimulatoryeffect, or no effect at all.

[0463] The amount and type of cytokine produced can be determined usingimmunological methods, such as ELISA assays. The methods of the presentinvention are not limited by the type of assay used to measure theamount of cytokine produced, and any methods known to those skilled inthe art and later developed can be used to measure the amount ofcytokines produced in response to the stimulating agent and to thecompound having unknown activity.

[0464] An aspect of the present invention comprises methods andcompositions for the treatment of diseases, preconditions or pathologiesassociated with inflammatory cytokines and other inflammation relatedmolecules including, but not limited to IL-6, VCAM-1, AGE-induced MCP-1,(monocyte chemoattractant protein 1), heme oxygenase, insulin-likegrowth factor, selecting, IP-10, MIG and I-TAC, NF-κB, IL-1β(interleukin 1β), IL-11 (interleukin 11), m-CSF (macrophage colonystimulating factor), fibrinogen, TNF-α (tumor necrosis factor α),adhesion molecules, selecting, VCAM-1 (Vascular Cell AdhesionMolecule-1), CRP (C-reactive protein), and PAI-1 (plasminogen activatorinhibitor-1). Examples of such diseases include the pathogenesis ofatherosclerosis and the development of diabetic vasculopathy in type IIdiabetes. For example, affecting the activity or level of TNFα is a keymediator of tissue damage following acute or chronic inflammatoryreactions. The present invention contemplates providing compositions andmethods that modulate the effects of cytokines and inflammatorymolecules such as TNFα, IL-6, VCAM-1, IP-10, MIG, I-TAC and AGE-inducedMCP-1, and treat the associated diseases, acute or chronic conditions,preconditions and pathologies.

[0465] Assays for determining the activity of compounds capable ofmodulating inflammation include those taught in U.S. patent applicationSer. Nos. 10/026,335 and 09/969,013, which are both expresslyincorporated by reference. In general, once the baseline response to thestimulating agent for the production of cytokines by the endothelialcells is established, thus comprising the control levels for thescreening assay, the methods comprise addition of compounds of thepresent invention. The effect of the compound on the baseline responseis determined by comparing the amount of cytokine produced in thepresence of the stimulating agent and the amount of cytokine produced inthe presence of the stimulating agent and the compound of the presentinvention. In a preferred method, compounds that have inhibitory effectson the inflammation of the cells in the presence of glycated albumin arethen used as therapeutic agents. One or more compounds may be added tothe screening assay. Combinations or mixtures of compounds can be added.Different amounts and formulations of the compounds are added todetermine the effects on the screening assay. The screening assay mayalso be used to determine stimulatory compounds or compounds that haveno effects in the assay.

[0466] The present invention comprises methods and compositions for thetreatment and prevention of disease, conditions and pathologiesassociated with inflammation. Such methods comprise administration ofcompositions comprising compounds capable of modulating the activity ofmolecules associated with inflammation such as AGE or cytokines or othercellular factors, including release rates or activity, and includecompositions comprising compounds disclosed herein with inflammationmodulating activity. Administration of such compounds that are effectivein modulating inflammation are administered to humans and animalssuspected of having or who have inflammatory diseases, for example,diabetic-induced vasculopathies, autoimmune diseases, renalinsufficiency, Alzheimer's syndrome, and inflammation-induced diseasessuch as atherosclerosis. Effective amounts are administered to suchhumans and animals in dosages that are safe and effective, including,but not limited to, the ranges taught herein. Routes of administrationinclude, but are not limited to, those disclosed herein. As disclosedherein, compositions comprising such compounds may be used inconjunction with other therapeutic agents or in methods comprising stepssuch as altered patient activities, including, but not limited to,changes in exercise or diet.

VI. Cytotoxic Activity

[0467] An embodiment of the present invention comprises methods andcompositions comprising compounds that have at least the activity ofcausing cellular death or a cessation of cellular activity, referred toherein as cytotoxic activity. This activity can be used in methods forin vitro or in vivo cytotoxicity. For example, compounds having thisactivity can be selectively delivered to an area within a livingorganism to selectively kill cells in that area. Such methods are usingin treating hyperproliferative cells, such as cancers, or other unwantedcellular growth or cellular activities. One aspect of the inventionprovides compositions comprising compounds that nonselectively killcells. Another aspect of the invention provides compounds thatselectively kill cells, for example, cells that have a particularcellular marker or other identifying characteristic such as metabolicrate or uptake of a particular compound, such as sodium, calcium orthymidine.

[0468] The present invention also provides compositions for and methodsof treatment of biological conditions including, but not limited to,conditions for which cytotoxic activity is a treatment. For example, thecompositions and methods for providing compounds that have at least theactivity of cytotoxicity are useful in the treatment or prophylaxis ofat least one hyperproliferative disease in a cell, tissue, organ,animal, or patient including, but not limited to, malignant andnon-malignant cell growth, leukemia, acute leukemia, acute lymphoblasticleukemia (ALL), B-cell, T-cell or FAB ALL, acute myeloid leukemia (AML),chromic myelocytic leukemia (CML), chronic lymphocytic leukemia (CLL),hairy cell leukemia, myelodyplastic syndrome (MDS), a lymphoma,Hodgkin's disease, a malignamt lymphoma, non-hodgkin's lymphoma,Burkitt's lymphoma, multiple myeloma, Kaposi's sarcoma, colorectalcarcinoma, pancreatic carcinoma, nasopharyngeal carcinoma, malignanthistiocytosis, paraneoplastic syndrome/hypercalcemia of malignancy,solid tumors, adenocarcinomas, sarcomas, malignant melanoma, hemangioma,metastatic disease, cancer related bone resorption, cancer related bonepain, and the like.

[0469] Compounds of the present invention that have at least theactivity of cytotoxicity are shown in TABLE 4A and B. The compoundsshown in this Table have the activity of cytotoxicty as measured by theassays taught herein. The inclusion of compounds in the categories ofthe Tables disclosed herein are not to be seen as limiting, in thatcompounds included in such Tables have at least the activity shown forinclusion in the Table and may have more or other activities. Nor arethe Tables to be seen as limiting in that these are the only compoundsdisclosed herein that have that activity, representative compounds areshown in the Tables that have at least that particular activity forinclusion in the Table. One or more compounds disclosed herein have atleast an activity that has utility in treatment of disease states.

[0470] Examples of compounds that show at least this activity andutility are shown in the following formula:

[0471] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0472] wherein:

[0473] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; cycloalkyl with up to 10carbon atoms; or aryl;

[0474] E is CH or N;

[0475] n is an integer from 0 to 3;

[0476] X¹ is selected from —H, m-F, m-Cl, m-Br, m-I, m-CN, m-NO₂,m-SO₂R¹, or m-SO₂OR¹, or X¹ and X² together is a fused benzene orpyridine ring;

[0477] X² is selected from —H, o-Cl, o-Br, p-OR¹, p-SR¹, p-NR¹ ₂,p-F,p-Cl, p-Br, p-CF₃, p-C(O)OR¹,p-OM, or p-SM, wherein M is selected fromLi, Na, K, Mg, or Ca;

[0478] A is selected from NR¹ or O, wherein Y¹ is selected fromcycloalkyl with up to 10 carbon atoms, linear or branched alkyl with upto 10 carbon atoms, or

[0479] when A is NR¹, and wherein Y¹ is selected from R¹ or CH₂R¹ when Ais O; or AY¹ is selected from a halogen,

[0480] and

[0481] DY² is a halogen, or D is NR¹ and y² is selected from

[0482] or (CHR¹)_(x)NR¹ ₂, wherein x is an integer from 1 to 6.

[0483] Additional examples of compounds that show at least this activityand utility are shown in the following formula:

[0484] or an ene, a diene, a triene, or an yne derivative thereof; asaturated derivative thereof; a stereoisomer thereof; or a salt thereof;

[0485] wherein:

[0486] R¹ is in each occurrence independently selected from —H; linearor branched alkyl with up to 10 carbon atoms; or cycloalkyl with up to10 carbon atoms;

[0487] X¹ is in each occurrence independently selected from —H, m-F,m-Cl, m-Br, m-I, m-CN, m-NO₂, m-SO₂R , or n-50₂OR¹;

[0488] X² is in each occurrence independently selected from o-CH₃,p-OR¹, p-SR¹, p-NR¹ ₂, or p-OM or p-SM, wherein M is selected from Li,Na, K, Mg, or Ca;

[0489] Y¹ is selected from cycloalkyl with up to 10 carbon atoms;

[0490] wherein n is 1 or 2; or

[0491] and

[0492] Y²is selected from

[0493] Further examples of compounds that show at least this activityand utility are presented in Tables 4A and 4B. The compound nomenclatureof Tables 4A and 4B, as in the other tables presented herein, wasgenerated using Autonom, where the name provided may be the Beulstein orCAS version of the chemical name. Note that any hydrogen atoms that arerequired for any atom to attain its usual valence in a structurepresented in Tables 4A and 4B, whether a carbon atom or a heteroatom,should be inferred if it is not specifically indicated. In addition,compounds, to include compositions thereof, encompassed within the scopeof structures Id-XIVd, respectively, or listed in Tables 1D and 1E maybe likewise employed in this embodiment and/or aspect of the presentinvention.

[0494] Assays for determining the activity of compounds capable ofcytotoxic activity include those taught in herein and others that arewell known in the art. In general, to determine if there is cytotoxicactivity associated with a compound, cells of a particular type, in agrowing stage or a quiescient stage, are treated with the compound ofinterest. Various parameters of cell death or cessation are used tomeasure the effects of the compound. For example, the amount of nucleicacid or protein synthesis can be measured or visual observation of thestate of the cells, such as release from the substrate, can be used tomeasure the state of the cells.

[0495] The present invention comprises methods and compositions for thetreatment and prevention of diseases or conditions that present orresult from cellular proliferation or unwanted cellular growth orcellular activity. Such methods comprise administration of compositionscomprising compounds capable of modulating cellular activity or causingcellular death or cessation of growth such as compositions comprisingcompounds disclosed herein that have cytotoxic activity. Administrationof such compounds that are effective in cytotoxic activity areadministered to humans and animals suspected of having or who have, forexample, cancer, overactive tissues such as thyroid or hypothalamus, orcellular conditions where factors are released in unwanted amounts.Effective amounts are administered to such humans and animals in dosagesthat are safe and effective, including, but not limited to, the rangestaught herein. Routes of administration include, but are not limited to,those disclosed herein. As disclosed herein, compositions comprisingsuch compounds may be used in conjunction with other therapeutic agentsor in methods comprising steps such as altered patient activities.

Compound/Composition-Coated Medical Devices

[0496] The compounds of the present invention may be used alone or incombination with other agents along with delivery devices to effectivelyprevent and treat the diseases described herein, though particularapplications are found in vascular disease, and in particular, vasculardisease caused by injury and/or by transplantation. Though this examplefocuses on vascular disease, provision of the compounds of the presentinvention with medical devices for treatment of the diseases andconditions capable of being treated with the compounds is contemplatedby the present invention.

[0497] Various medical treatment devices utilized in the treatment ofvascular disease may ultimately induce further complications. Forexample, balloon angioplasty is a procedure utilized to increase bloodflow through an artery and is the predominant treatment for coronaryvessel stenosis. However, the procedure typically causes a certaindegree of damage to the vessel wall, thereby creating new problems orexacerbating the original problem at a point later in time. Althoughother procedures and diseases may cause similar injury, exemplaryembodiments of the present invention will be described with respect tothe treatment of restenosis and related complications followingpercutaneous transluminal coronary angioplasty and other similararterial/venous procedures, including the joining of arteries, veins andother fluid carrying conduits in other organs or sites of the body, suchas the liver, lung, bladder, kidney, brain, prostate, neck and legs.

[0498] The local delivery of a compound of the present invention and, insome embodiments, along with other therapeutic agents, from a stentprevents vessel recoil and remodeling through the scaffolding action ofthe stent. The activity of compound provided, with or without othertherapeutic agents, helps determine for which application, to treatwhich disease, the coated medical device is being administered. Forexample, compound-coated stents can prevent multiple components ofneointimal hyperplasia or restenosis as well as reduce inflammation andthrombosis. Local administration of a compound of the present inventionand other therapeutic agents to stented coronary arteries may also haveadditional therapeutic benefit. For example, higher tissueconcentrations of the compounds of the present invention and othertherapeutic agents may be achieved utilizing local delivery rather thansystemic administration. In addition, reduced systemic toxicity may beachieved utilizing local delivery rather than systemic administrationwhile maintaining higher tissue concentrations. In utilizing localdelivery from a stent rather than systemic administration, a singleprocedure may suffice with better patient compliance. An additionalbenefit of combination therapeutic agent and/or compound therapy may beto reduce the dose of each of the therapeutic agents, thereby limitingtoxicity, while still achieving a reduction in restenosis, inflammationand thrombosis. Local stent-based therapy is therefore a means ofimproving the therapeutic ratio (efficacy/toxicity) of anti-restenosis,anti-inflammatory, and anti-thrombotic therapeutic agents.

[0499] Although exemplary embodiments of the invention will be describedwith respect to the treatment of restenosis and other relatedcomplications, it is important to note that the local delivery of acompound of the present invention, alone or as part of a therapeuticagent combination, may be utilized to treat a wide variety of conditionsutilizing any number of medical devices, or to enhance the functionand/or life of the device. For example, intraocular lenses, placed torestore vision after cataract surgery is often compromised by theformation of a secondary cataract. The latter is often a result ofcellular overgrowth on the lens surface and can be potentially minimizedby combining one or more compounds of the present invention havingactivity that is effecting in proventing unwanted cellular growth withthe device. Other medical devices that often fail due to tissuein-growth or accumulation of proteinaceous material in, on and aroundthe device, such as shunts for hydrocephalus, dialysis grafts, colostomybag attachment devices, ear drainage tubes, leads for pace makers andimplantable defibrillators can also benefit from the combinations of thecompounds of the present invention, possibly other pharmaceuticalagents, and the devices. Other surgical devices, sutures, staples,anastornosis devices, vertebral disks, bone pins, suture anchors,hemostatic barriers, clamps, screws, plates, clips, vascular implants,tissue adhesives and sealants, tissue scaffolds, various types ofdressings, bone substitutes, intraluminal devices, and vascular supportscould also provide enhanced patient benefit using this compound-devicecombination approach. Essentially, any type of medical device may becoated in some fashion with at least one compound of the presentinvention, alone or as part of a therapeutic agent combination, whichenhances treatment over the use of the device or therapeutic agentwithout combination with the compound.

[0500] As disclosed supra, the compounds of the present invention can beadministered in combinational therapies with other therapeutic agents,and are not limited to only the other therapeutic agents disclosedherein. Thus, the present invention also contemplates, in addition tovarious medical devices, the coatings on these devices may be used todeliver a compound of the present invention in combination with othertherapeutic agents. This illustrative list of therapeutic agents can beadministered through pharmeutical means or in association with medicaldevices and such therapeutic agents include, but are not limited to,antiproliferative/antimitotic agents including natural products such asvinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine),paclitaxel, epidipodophyllotoxins (e.g., etoposide, teniposide),antibiotics (dactinomycin (actinomycin D) daunorubicin, doxorubicin andidarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin(mithramycin) and mitomycin, enzymes (L-asparaginase which systemicallymetabolizes L-asparagine and deprives cells which do not have thecapacity to synthesize their own asparagine); antiplatelet agents suchas G(GP) IIb/IIIa inhibitors and vitronectin receptor antagonists;antiproliferative/antimitotic alkylating agents such as nitrogenmustards (mechlorethamine, cyclophosphamide and analogs, melphalan,chlorambucil), ethylenimines and methylmelamines (hexamethylmelamine andthiotepa), alkyl sulfonates-busulfan, nirtosoureas (carmustine (BCNU)and analogs, streptozocin), trazenes-dacarbazinine (DTIC);antiproliferative/antimitotic antimetabolites such as folic acid analogs(methotrexate), pyrimidine analogs (fluorouracil, floxuridine, andcytarabine), purine analogs and related inhibitors (mercaptopurine,thioguanine, pentostatin and 2-chlorodeoxyadenosine (cladribine));platinum coordination complexes (cisplatin, carboplatin), procarbazine,hydroxyurea, mitotane, aminoglutethimide; hormones (e.g. estrogen);anticoagulants (heparin, synthetic heparin salts and other inhibitors ofthrombin); fibrinolytic agents (such as tissue plasminogen activator,streptokinase and urokinase), aspirin, dipyridamole, ticlopidine,clopidogrel, abeiximab; antimigratory; antisecretory (breveldin);anti-inflammatory agents such as adrenocortical steroids (cortisol,cortisone, fludrocortisone, prednisone, prednisolone,6α-methylprednisolone, triamcinolone, betamethasone, and dexamethasone),non-steroidal agents (salicylic acid derivatives, i.e., aspirin;para-aminophenol derivatives, i.e., acetominophen; indole and indeneacetic acids (indomethacin, sulindac, and etodalac), beteroaryl aceticacids (tolmetin, diclofenac, and ketorolac), arylpropionic acids(ibuprofen and derivatives), anthranilic acids (mefenamic acid, andmeclofenamic acid), enolic acids (piroxicam, tenoxicam, phenylbutazone,and oxyphenthatrazone), nabumetone, gold compounds (auranofin,aurothioglucose, gold sodium thiomalate); immunosuppressives.(Cyclosporine, tacrolimus (FK-506), sirolimus (rapamycin), azathioprine,mycophenolate mofetil); angiogenic agents: vascular endothelial growthfactor (VEGF), fibroblast growth factor (FGF); angiotensin receptorblockers; nitric oxide donors; anti-sense oligionucleotides andcombinations thereof; cell cycle inhibitors, mTOR inhibitors, and growthfactor signal transduction kinase inhibitors.

[0501] Although any number of stents may be utilized in accordance withthe present invention, for simplicity, a limited number of stents willbe described in exemplary embodiments of the present invention. Theskilled artisan will recognize that any number of stents may be utilizedin connection with the present invention. In addition, as stated above,other medical devices may be utilized. For example, though stents aredescribed, sleeves outside the vessels are also contemplated, as areother medical devices that can provide a substrate for administrationfor at least one of the compounds of the present invention.

[0502] A stent is commonly used as a tubular structure left inside thelumen of a duct to relieve an obstruction. Typically, stents areinserted into the lumen in a non-expanded form and are then expandedautonomously, or with the aid of a second device in situ. A commonmethod of expansion occurs through the use of a catheter-mounted,angioplasty balloon that is inflated within the stenosed vessel or bodypassageway in order to shear and disrupt the obstructions associatedwith the wall components of the vessel and to obtain an enlarged lumen.

[0503] A stent may resemble an expandable cylinder and may comprise afenestrated structure for placement in a blood vessel, duct or lumen tohold the vessel, duct or lumen open, more particularly for protecting asegment of artery from restenosis after angioplasty. The stent may beexpanded circumferentially and maintained in an expanded configurationthat is circumferentially or radially rigid. The stent may be axiallyflexible and when flexed at a band, for example, the stent avoids anyexternally protruding component parts.

[0504] The stent may be fabricated utilizing any number of methods. Forexample, the stent may be fabricated from a hollow or formed stainlesssteel tube that may be machined using lasers, electric dischargemilling, chemical etching or other means. The stent is inserted into thebody and placed at the desired site in an unexpanded form. In oneembodiment, expansion may be effected in a blood vessel by a ballooncatheter, where the final diameter of the stent is a function of thediameter of the balloon catheter used. It should be appreciated that astent in accordance with the present invention may be embodied in ashape-memory material including, for example, an appropriate alloy ofnickel and titanium or stainless steel.

[0505] Structures formed from stainless steel may be made self-expandingby configuring the stainless steel in a predetermined manner, forexample, by twisting it into a braided configuration. In thisembodiment, after the stent has been formed it may be compressed so asto occupy a space sufficiently small as to permit its insertion in ablood vessel or other tissue by insertion means, wherein the insertionmeans include a suitable catheter, or flexible rod. Upon emerging fromthe catheter, the stent may be configured to expand into the desiredconfiguration where the expansion is automatic or triggered by a changein pressure, temperature or electrical stimulation.

[0506] Furthermore, a stent may be modified to comprise one or morereservoirs. Each of the reservoirs may be opened or closed as desired.These reservoirs may be specifically designed to hold the the compoundor compound/therapeutic agent combination to be delivered. Regardless ofthe design of the stent, it is preferable to have the compound orcompound/therapeutic agent combination dosage applied with enoughspecificity and a sufficient concentration to provide an effectivedosage in the affected area. In this regard, the reservoir size in thebands is preferably sized to adequately apply the the compound orcompound/therapeutic agent combination dosage at the desired locationand in the desired amount.

[0507] In an alternative embodiment, the entire inner and outer surfaceof the stent may be coated with the compound or compound/therapeuticagent combination in therapeutic dosage amounts. The coating techniquesmay vary depending on the the compound or compound/therapeutic agentcombination. Also, the coating techniques may vary depending on thematerial comprising the stent or other intraluminal medical device.

[0508] One or more compounds of the present invention and, in someinstances, other therapeutic agents as a combination, may beincorporated onto or affixed to the stent in a number of ways. In oneembodiment, the compound is directly incorporated into a polymericmatrix and sprayed onto the outer surface of the stent. The compoundelutes from the polymeric matrix over time and enters the surroundingtissue. The compound preferably remains on the stent for at least threedays up to approximately six months, and more preferably between sevenand thirty days.

[0509] Any number of non-erodible polymers may be utilized inconjunction with the compound, and such polyermic compositions are wellknown in the art. In one embodiment, the polymeric matrix comprises twolayers. The base layer comprises a solution ofpoly(ethylene-covinylacetate) and polybutylmethacrylate. The compound isincorporated into this base layer. The outer layer comprises onlypolybutylmethacrylate and acts as a diffusion barrier to prevent thecompound from eluting too quickly. The thickness of the outer layer ortopcoat determines the rate at which the compound elutes from thematrix. Essentially, the compound elutes from the matrix by diffusionthrough the polymer matrix. Polymers are permeable, thereby allowingsolids, liquids and gases to escape therefrom. The total thickness ofthe polymeric matrix is in the range from about one micron to abouttwenty microns or greater. It is important to note that primer layersand metal surface treatments may be utilized before the polymeric matrixis affixed to the medical device. For example, acid cleaning, alkaline(base) cleaning, salinization and parylene deposition may be used aspart of the overall process described above.

[0510] The poly(ethylene-co-vinylacetate), polybutylmethacrylate andcompound solution may be incorporated into or onto the stent in a numberof ways. For example, the solution may be sprayed onto the stent or thestent may be dipped into the solution. Other methods include spincoating and plasma polymerization. In one embodiment, the solution issprayed onto the stent and then allowed to dry. In another embodiment,the solution may be electrically charged to one polarity and the stentelectrically charged to the opposite polarity. In this manner, thesolution and stent will be attracted to one another. In using this typeof spraying process, waste may be reduced and more precise control overthe thickness of the coat may be achieved.

[0511] Drug-coated stents are manufactured by a number of companiesincluding Johnson & Johnson, Inc. (New Brunswick, N.J.), Guidant Corp.(Santa Clara, Calif.), Medtronic, Inc. (Minneapolis, Minn.), Cook GroupIncorporated (Bloomington, Ind.), Abbott Labs., Inc. (Abbott Park,Ill.), and Boston Scientific Corp. (Natick, Mass.). See e.g., U.S. Pat.No. 6,273, 913; U.S. patent application Ser. No. 2002/0051730; WO02/26271; and WO 02/26139, each expressly entirely incorporated hereinby reference.

Expression Profiles and Microarray Methods of Use

[0512] Other aspects of the present invention comprise compositions andmethods for microarray devices. Such microarray devices and methodscomprise a variety of microarrays that may be used, for example, tostudy and monitor gene expression in response to treatment with thecompounds of the present invention. The microarrays may comprise nucleicacid sequences, carbohydrates or proteins that are determinative forspecific cells, tissues, species, disease states, prognoses, diseaseprogression, or any other combination of molecules that can be used todetermine an effect of one or more of the compounds of the presentinvention

[0513] For example, the microarrays of the present invention may bederived from, or representative of, for example, a specific organism orcell type, including human microarrays, vascular microarrays,inflammation microarrays, cancer microarrays, apoptosis microarrays,oncogene and tumor suppressor microarrays, cell-cell interactionmicroarrays, cytokine and cytokine receptor microarrays, bloodmicroarrays, cell cycle microarrays, neuroarrays, mouse microarrays, andrat microarrays, or combinations thereof. In further embodiments, themicroarrays may represent diseases including cardiovascular diseases,vasculopathic conditions, inflammatory diseases, autoimmune diseases,neurological diseases, immunological diseases, various cancers,infectious diseases, endocrine disorders, and genetic diseases.

[0514] Alternatively, the microarrays useful in assessing the efficacyof the compounds of the present invention may represent a particulartissue type including, but not limited to, heart, liver, prostate, lung,nerve, muscle, or connective tissue; preferably coronary arteryendothelium, umbilical artery endothelium, umbilical vein endothelium,aortic endothelium, dermal microvascular endothelium, pulmonary arteryendothelium, myometrium microvascular endothelium, keratinocyteepithelium, bronchial epithelium, mammary epithelium, prostateepithelium, renal cortical epithelium, renal proximal tubule epithelium,small airway epithelium, renal epithelium, umbilical artery smoothmuscle, neonatal dermal fibroblast, pulmonary artery smooth muscle,dermal fibroblast, neural progenitor cells, skeletal muscle, astrocytes,aortic smooth muscle, mesangial cells, coronary artery smooth muscle,bronchial smooth muscle, uterine smooth muscle, lung fibroblast,osteoblasts, prostate stromal cells, or combinations thereof.

[0515] The present invention further contemplates microarrays comprisinga gene expression profile comprising one or more polynucleotidesequences including complementary and homologous sequences, wherein saidgene expression profile is generated from a cell type treated with acompound of the present invention and is selected from the groupcomprising coronary artery endothelium, umbilical artery endothelium,umbilical vein endothelium, aortic endothelium, dermal microvascularendothelium, pulmonary artery endothelium, myometrium microvascularendothelium, keratinocyte epithelium, bronchial epithelium, mammaryepithelium, prostate epithelium, lenal cortical epithelium, renalproximal tubule epithelium, small airway epithelium, renal epithelium,umbilical artery smooth muscle, neonatal dermal fibroblast, pulmonaryartery smooth muscle, dermal fibroblast, neural progenitor cells,skeletal muscle, astrocytes, aortic smooth muscle, mesangial cells,coronary artery smooth muscle, bronchial smooth muscle, uterine smoothmuscle, lung fibroblast, osteoblasts, and prostate stromal cells.

[0516] The present invention contemplates microarrays comprising one ormore protein-binding agents, wherein a protein expression profile isgenerated from a cell type treated with a compound of the presentinvention and is selected from the group comprising coronary arteryendothelium, umbilical artery endothelium, umbilical vein endothelium,aortic endothelium, dermal microvascular endothelium, pulmonary arteryendothelium, myometrium microvascular endothelium, keratinocyteepithelium, bronchial epithelium, mammary epithelium, prostateepithelium, renal cortical epithelium, renal proximal tubule epithelium,small airway epithelium, renal epithelium, umbilical artery smoothmuscle, neonatal dermal fibroblast, pulmonary artery smooth muscle,dermal fibroblast, neural progenitor cells, skeletal muscle, astrocytes,aortic smooth muscle, mesangial cells, coronary artery smooth muscle,bronchial smooth muscle, uterine smooth muscle, lung fibroblast,osteoblasts, and prostate stromal cells.

[0517] More specifically, the present invention contemplates methods forthe reproducible measurement and assessment of the expression ofspecific mRNAs or proteins in, for example, a specific set of cells. Onemethod combines and utilizes the techniques of laser capturemicrodissection, T7-based RNA amplification, production of cDNA fromamplified RNA, and DNA microarrays containing immobilized DNA moleculesfor a wide variety of specific genes, including HSPGs such as perlecan,to produce a profile of gene expression analysis for very small numbersof specific cells. The desired cells are individually identified andattached to a substrate by the laser capture technique, and the capturedcells are then separated from the remaining cells. RNA is then extractedfrom the captured cells and amplified about one million-fold using theT7-based amplification technique, and cDNA may be prepared from theamplified RNA. A wide variety of specific DNA molecules are preparedthat hybridize with specific polynucleotides of the microarray, and theDNA molecules are immobilized on a suitable substrate. The cDNA madefrom the captured cells is applied to the microarray under conditionsthat allow hybridization of the cDNA to the immobilized DNA on themicroarray. The expression profile of the captured cells is obtainedfrom the analysis of the hybridization results using the amplified RNAor cDNA made from the amplified RNA of the captured cells, and thespecific immobilized DNA molecules on the microarray. The hybridizationresults demonstrate, for example, which genes of those represented onthe microarray as probes are hybridized to cDNA from the captured cells,and/or the amount of specific gene expression. The hybridization resultsrepresent the gene expression profile of the captured cells. The geneexpression profile of the captured cells can be used to compare the geneexpression profile of a different set of captured cells. For example,gene expression profiles may be generated from cells treated (and nottreated) with a compound of the present invention. The similarities anddifferences provide useful information for determining the differencesbetween the same cell type under different conditions, morespecifically, the change in gene expression in response to treatmentwith a compound of the present invention.

[0518] The techniques used for gene expression analysis are likewiseapplicable in the context of protein expression profiles. Total proteinmay be isolated from a cell sample and hybridized to a microarraycomprising a plurality of protein-binding agents, which may includeantibodies, receptor proteins, small molecules, and the like. Using anyof several assays known in the art, hybridization may be detected andanalyzed as described above. In the case of fluorescent detection,algorithms may be used to extract a protein expression profilerepresentative of the particular cell type. In this regard, the changein protein expression in response to treatment of cells with a compoundof the present invention may be evaluated.

[0519] Thus, in one aspect, the present invention comprises at least onemicroarray corresponding to a population of genes isolated from aparticular tissue or cell type in methods that is used to detect changesin gene transcription levels that result from exposing the selectedtissue or cells to at least one compound of the present invention. Inthis embodiment, a biological sample derived from an organism, or anestablished cell line, may be exposed to at least one compound of thepresent invention in vivo or ex vivo. Thereafter, the gene transcripts,primarily mRNA, of the tissue or cells are isolated by methodswell-known in the art. SAMBROOK ET AL., MOLECULAR CLONING: A LAB. MANUAL(2001). The isolated transcripts are then contacted with a microarrayunder conditions where the transcripts hybridize with a correspondingprobe to form hybridization pairs. Thus, the microarray provides a modelof the transcriptional responsiveness following exposure to at least onecompound of the present invention. Such information can be used todetermine therapeutic candidates. A hybridization signal may then bedetected at each hybridization pair to obtain a gene expression profile.

[0520] Gene and/or protein expression profiles and microarrays may alsobe used to identify activating or non-activating compounds of aparticular gene such as perlecan or other HSPG. Compounds that increasetranscription rates or stimulate, maintain, or stabilize the activity ofa protein are considered activating, and compounds that decrease ratesor inhibit the activity of a protein are non-activating. Moreover, thebiological effects of a compound may be reflected in the biologicalstate of a cell. This state is characterized by the cellularconstituents. One aspect of the biological state of a cell is itstranscriptional state. The transcriptional state of a cell includes theidentities and amounts of the constituent RNA species, especially mRNAs,in the cell under a given set of conditions. Thus, the gene expressionprofiles, microarrays, and algorithms discussed herein may be used toanalyze and characterize the transcriptional state of a given cell ortissue following exposure to an activating or non-activating compound,specifically, a compound of the present invention.

[0521] Microarray techniques and methods for analyzing results are wellknown in the art. See U.S. Pat. Nos. 6,263,287; 6,239,209; 6,218,122;6,197,599; 6,156,501; 5,874,219; 5,837,832; 5,700,637; 5,445,934; U.S.patent application Nos. 2001/0014461 A1; 2001/0039016 A1; 2001/0034023A1; WO 01/94946; and WO 01/77668. See also, Haab et al., 2 GENOMEBIOLOGY 1-12 (2001); Brown et al., 97 PROC. NATL. ACAD. SCI. USA 262-7(2000); Getz et al., 97 PROC. NATL. ACAD. SCL. USA 12079-84 (2000);Harrington et al., 3 CURRENT OPINION MICROBIOL 285-91 (2000); Holter etal., 97 PROC. NATL. ACAD. SCI. USA 8409-14 (2000); MacBeath et al., 289SCIENCE 1760-63 (2000); Duggan et al., 21 NATURE GENET 10-14 (1999);Lipshutz et al., 21 NATURE GENET 5-9 (1999); Eisen et al., 95 PROC.NATL. ACAD. SCI. USA 14863-68 (1998); Ermolaeva et al., 20 NATURE GENET.19-23 (1998); Hacia et al., 26 NUCLEIC ACIDS RES. 3865-66 (1998);Lockhart et al., NUCLEIC ACIDS SYMP. SER. 11-12 (1998); Schena et al.,16 TRENDS BIOTECHNOL. 301-6 (1998); Shalon, 46 PATHOL. BIOL. 107-9(1998); Welford et al., 26 NUCLEIC ACID RES. 3059-65 (1998); Blanchardet al., 11 BIOSENSORS BIOELECTRONICS 687-90 (1996); Lockhart et al., 14NATURE BIOTECHNOL. 1675-80 (1996); Schena et al., 93 PROC. NATL. ACAD.SCI. USA 10614-19 (1996); Tomayo et al., 96 PROC. NATL. ACAD. SCI. USA2907-12 (1996); Schena et al., 270 SCIENCE 467-70 (1995)

[0522] Database Creation, Database Access and Associated Methods of Use

[0523] Another embodiment of the present invention comprises a varietyof methods for managing or using data related to the compounds, methodsof making the compounds, methods of using and administering thecompounds, and diagnosing, prognosing and following the outcomesassociated with diseases in which the compounds are effective intreating. For example, methods for providing diagnostics and predictorsrelating to biomolecules including HSPGS, particularly, perlecan, arecontemplated by the present invention. Also within the scope of thisinvention are methods providing diagnostics and predictors relating tothe efficacy of the compounds of the present invention. The presentinvention further contemplates methods of providing expression profiledatabases, and methods for producing such databases, for normal anddiseased tissues.

[0524] The expression profile database may be an internal databasedesigned to include annotation information about the expression profilesgenerated to assess the effect of the compounds of the present inventionand through other sources and methods. Such information may include, forexample, the databases in which a given biomolecule was found, patientinformation associated with the expression profile, including age,cancer or tumor type or progression, information related to a compoundof the present invention such as dosage and administration information,descriptive information about related cDNAs associated with thesequence, tissue or cell source, sequence data obtained from externalsources, expression profiles for a given gene and the related diseasestate or course of disease, for example whether the expression profilerelates to or signifies a particular disease state, and preparationmethods. The expression profiles may be based on protein and/orpolynucleotide microarray data obtained from publicly available orproprietary sources. The database may be divided into two sections: onefor storing the sequences and related expression profiles and the otherfor storing the associated information. This database may be maintainedas a private database with a firewall within the central computerfacility. However, this invention is not so limited and the expressionprofile database may be made available to the public.

[0525] The database may be a network system connecting the networkserver with clients. The network may be any one of a number ofconventional network systems, including a local area network (LAN) or awide area network (WAN), as is known in the art (e.g., Ethernet). Theserver may include software to access database information forprocessing user requests, and to provide an interface for servinginformation to client machines. The server may support the World WideWeb and maintain a website and Web browser for client use. Client/serverenvironments, database servers, and networks are well documented in thetechnical, trade, and patent literature.

[0526] Through the Web browser, clients may construct search requestsfor retrieving data from, for example, a microarray database and anexpression profile database. For example, the user may “point and click”to user interface elements such as buttons, pull down menus, and scrollbars. The client requests may be transmitted to a Web application thatformats them to produce a query that may be used to gather informationfrom the system database, based, for example, on microarray orexpression data obtained by the client, and/or other phenotypic orgenotypic information. Specifically, the client may submit expressiondata based on microarray expression profiles obtained from a patienttreated with a compound of the present invention and use the system toobtain a diagnosis based on that information based on a comparison bythe system of the client expression data with the expression datacontained in the database. By way of example, the system compares theexpression profiles submitted by the client with expression profilescontained in the database and then provides the client with diagnosticinformation based on the best match of the client expression profileswith the database profiles. Thus, in one aspect, the comparison ofexpression profiles aids the clinician in determining the effectivenessof treatment with a compound of the present invention. Based on such acomparison, the clinician may alter or adjust the treatment regimen.

[0527] In addition, the website may provide hypertext links to publicdatabases such as GenBank and associated databases maintained by theNational Center for Biotechnology Information (NCBI), part of theNational Library of Medicine as well as, any links providing relevantinformation for gene expression analysis, genetic disorders, scientificliterature, and the like. Information including, but not limited to,identifiers, identifier types, biomolecular sequences, common clusteridentifiers (GenBank, Unigene, Incyte template identifiers, and soforth) and species names associated with each gene, is contemplated.

[0528] The present invention also provides a system for accessing andcomparing bioinformation, specifically expression profiles and otherinformation which is useful in the context of the compositions andmethods of the present invention. In one embodiment, the computer systemmay comprise a computer processor, suitable memory that is operativelycoupled to the computer processor, and a computer process stored in thememory that executes in the computer processor and which comprises ameans for matching an expression profile of a biomolecular sequence froma patient with expression profile and sequence identificationinformation of biomolecular sequences in a database. More specifically,the computer system is used to match an expression profile generatedfrom a biological sample treated with a compound of the presentinvention with expression profile and other information in a database.

[0529] Furthermore, the system for accessing and comparing informationcontained in biomolecular databases comprises a computer programcomprising computer code providing an algorithm for matching anexpression profile generated from a patient, for example, treated with acompound of the present invention, with expression profile and sequenceidentification information of biomolecular sequences in a biomoleculardatabase.

[0530] The present invention contemplates, in one embodiment, the use ofa Graphical User Interface (“GUI”) for the access of expression profileinformation stored in a biomolecular database. In a specific embodiment,the GUI may be composed of two frames. A first frame may contain aselectable list of biomolecular databases accessible by the user. When abiomolecular database is selected in the first frame, a second frame maydisplay information resulting from the pair-wise comparison of theexpression profile database with the client-supplied expression profileas described above, along with any other phenotypic or genotypicinformation.

[0531] The second frame of the GUI may contain a listing of biomolecularsequence expression information and profiles contained in the selecteddatabase. Furthermore, the second frame may allow the user to select asubset, including all of the biomolecular sequences, and to perform anoperation on the list of biomolecular sequences. In one embodiment, theuser may select the subset of biomolecular sequences by selecting aselection box associated with each biomolecular sequence. In anotherembodiment, the operations that may be performed include, but are notlimited to, downloading all listed biomolecular sequences to a databasespreadsheet with classification information, saving the selected subsetof biomolecular sequences to a user file, downloading all listedbiomolecular sequences to a database spreadsheet without classificationinformation, and displaying classification information on a selectedsubset of biomolecular sequences.

[0532] If the user chooses to display classification information on aselected subset of biomolecular sequences, a second GUI may be presentedto the user. In one embodiment, the second GUI may contain a listing ofone or more external databases used to create the expression profiledatabases as described above. Furthermore, for each external database,the GUI may display a list of one or more fields associated with eachexternal database. In yet another embodiment, the GUI may allow the userto select or deselect each of the one or more fields displayed in thesecond GUI. In yet another embodiment, the GUI may allow the user toselect or deselect each of the one or more external databases.

[0533] The methods of the present application further relate to thecommercial and other uses of the compositions and methodologies of thepresent invention. In one aspect, the methods include the marketing,sale, or licensing of the compositions and methodologies of the presentinvention in the context of providing consumers, i.e., patients, medicalpractitioners, medical service providers, researchers, andpharmaceutical distributors and manufacturers, with expression profiledatabases including, in particular, databases produced in accordancewith the use of the compounds of the present invention.

[0534] In another embodiment, the methods of the present inventioninclude establishing a distribution system for distributing thepharmaceutical compositions of the present invention for sale, and mayoptionally include establishing a sales group for marketing thepharmaceutical composition. Yet another aspect of the present inventionprovides a method of conducting target discovery comprising identifying,by one or more of the above drug discovery methods, a test compound, asdescribed above, which modulates the level of expression of a gene orthe activity of a gene product such as perlecan; conducting therapeuticprofiling of agents identified, or further analogs thereof, for efficacyand toxicity in animals; and optionally formulating a pharmaceuticalcomposition including one or more of the agents identified as having anacceptable therapeutic profile; and optionally licensing or selling, therights for further drug development of said identified agents.

[0535] Pharmaceutical Compositions

[0536] In addition to the compounds disclosed herein, the pharmaceuticalcompositions of the present invention can further comprise at least oneof any suitable auxiliary such as, but not limited to, diluent, binder,stabilizer, buffers, salts, lipophilic solvents, preservative, adjuvantor the like. Pharmaceutically acceptable auxiliaries are preferred.Examples and methods of preparing such sterile solutions are well knownin the art and can be found in well known texts such as, but not limitedto, REMINGTON'S PHARMACEUTICAL SCIENCES (Gennaro, Ed., 18th Edition,Mack Publishing Co. (1990)). Pharmaceutically acceptable carriers can beroutinely selected that are suitable for the mode of administration,solubility and/or stability of the compound.

[0537] Pharmaceutical excipients and additives useful in the presentinvention include, but are not limited to, proteins, peptides, aminoacids, lipids, and carbohydrates (e.g., sugars, includingmonosaccharides, di-, tri-, tetra-, and oligosaccharides; derivatizedsugars such as alditols, aldonic acids, esterified sugars and the like;and polysaccharides or sugar polymers), which can be present singly orin combination, comprising alone or in combination in ranges of 1-99.99%by weight or volume. Exemplary protein excipients include serum albuminsuch as human serum albumin (HSA), recombinant human albumin (rHA),gelatin, casein, and the like. Representative amino acid components,which can also function in a buffering capacity, include alanine,glycine, arginine, betaine, histidine, glutamic acid, aspartic acid,cysteine, lysine, leucine, isoleucine, valine, methionine,phenylalanine, aspartame, and the like.

[0538] Carbohydrate excipients suitable for use in the present inventioninclude, for example, monosaccharides such as fructose, maltose,galactose, glucose, D-mannose, sorbose, and the like; disaccharides,such as lactose, sucrose, trehalose, cellobiose, and the like;polysaccharides, such as raffinose, melezitose, maltodextrins, dextrans,starches, and the like; and alditols, such as mannitol, xylitol,maltitol, lactitol, xylitol, sorbitol (glucitol), myoinositol and thelike.

[0539] The pharmaceutical compositions comprising the compounds of thepresent invention can also include a buffer or a pH adjusting agent.Typically, the buffer is a salt prepared from an organic acid or base.Representative buffers include organic acid salts such as salts ofcitric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid,succinic acid, acetic acid, or phthalic acid; Tris, tromethaminehydrochloride, or phosphate buffers.

[0540] Additionally, pharmaceutical compositions of the invention caninclude polymeric excipients/additives such as polyvinylpyrrolidones,ficolls (a polymeric sugar), dextrates (e.g., cyclodextrins, such as2-hydroxypropyl-β-cyclodextrin), polyethylene glycols, flavoring agents,anti-microbial agents, sweeteners, antioxidants, anti-static agents,surfactants (e.g., polysorbates such as “TWEEN 20” and “TWEEN 80”),lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol),and chelating agents (e.g., EDTA). These and additional knownpharmaceutical excipients and/or additives suitable for use in thepresent invention are known in the art, e.g., as listed in REMINGTON:THE SCIENCE & PRACTICE OF PHARMACY (19^(th) ed., Williams & Williams(1995)) and PHYSICIAN'S DESK REFERENCE (52^(nd) ed., Medical Economics(1998)), the disclosures of which are expressly entirely incorporatedherein by reference.

[0541] Pharmaceutical Compositions for Oral Administration

[0542] For oral administration in the form of a tablet or capsule, acompound may be combined with an oral, non-toxic pharmaceuticallyacceptable inert carrier such as ethanol, glycerol, water and the like.Moreover, when desired or necessary, suitable binders, lubricants,disintegrating agents, and coloring agents may also be incorporated intothe mixture. Suitable binders include, without limitation, starch;gelatin; natural sugars such as glucose or beta-lactose; cornsweeteners; natural and synthetic gums such as acacia, tragacanth, orsodium alginate, carboxymethylcellulose; polyethylene glycol; waxes andthe like. Lubricants used in these dosage forms include, withoutlimitation, sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride and the like. Disintegratorsinclude, without limitation, starch, methyl cellulose, agar, bentonite,xanthan gum and the like.

[0543] Formulations of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil emulsion and as a bolus, etc.

[0544] A tablet may be made by compression or molding, optionally withone or more accessory ingredients. Compressed tablets may be prepared bycompressing, in a suitable machine, the active ingredient in afree-flowing form such as a powder or granules, optionally mixed with abinder, lubricant, inert diluent, preservative, surface active ordispersing agent. Molded tablets may be made by molding, in a suitablemachine, a mixture of the powdered compound moistened with an inertliquid diluent. The tablets may be optionally coated or scored and maybe formulated so as to provide a slow or controlled release of theactive ingredient therein.

[0545] In addition, the combinations may be incorporated intobiodegradable polymers allowing for sustained release of the compound,the polymers being implanted in the vicinity of where drug delivery isdesired, for example, at the site of restenosis. The biodegradablepolymers and their uses are described, for example, in detail in Brem etal., 74 J. NEUROSURG. 441-46 (1991). Suitable examples ofsustained-release compositions include semipermeable matrices of solidhydrophobic polymers containing a compound of the present invention,which matrices are in the form of shaped articles, e.g., films, ormicrocapsules. Examples of sustained-release matrices includepolyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate),or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919),copolymers of L-glutamic acid and y ethyl-L-glutamate, non-degradableethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymerssuch as the LUPRON DEPOT® (Tap Pharmaceuticals, Inc., Chicago, Ill.)(injectable microspheres composed of lactic acid glycolic acid copolymerand leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid.

[0546] Pharmaceutical Compositions for Parenteral Administration

[0547] Formulations suitable for parenteral administration includeaqueous and non-aqueous sterile injection solutions which may containanti-oxidants, buffers, bacteriostats and solutes that render theformulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example, sealed ampules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for example,water for injections, immediately prior to use. Extemporaneous injectionsolutions and suspensions may be prepared from sterile powders, granulesand tablets of the kind previously described.

[0548] For parenteral administration, sterile suspensions and solutionsare desired. Isotonic preparations which generally contain suitablepreservatives are employed when intravenous administration is desired.The pharmaceutical compositions may be administered parenterally viainjection of a formulation consisting of the active ingredient dissolvedin an inert liquid carrier. The term “parenteral,” as used herein,includes, but is not limited to, subcutaneous injections, intravenous,intramuscular, intraperitoneal injections, or infusion techniques.Acceptable liquid carriers include, for example, vegetable oils such aspeanut oil, cotton seed oil, sesame oil and the like, as well as organicsolvents such as solketal, glycerol formal and the like. Theformulations may be prepared by dissolving or suspending the activeingredient in the liquid carrier such that the final formulationcontains from about 0.005% to 30% by weight of the active ingredient,i.e., a compound of the present invention.

[0549] Pharmaceutical Compositions for Other Routes of Administration

[0550] Formulations suitable for topical administration in the mouthinclude lozenges comprising the ingredients in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the compound to be administeredin a suitable liquid carrier. The liquid forms may include suitablyflavored suspending or dispersing agents such as the synthetic andnatural gums, for example, tragacanth, acacia, methyl-cellulose and thelike.

[0551] Formulations for rectal administration may be presented as asuppository with a suitable base comprising, for example, cocoa butteror a salicylate.

[0552] Formulations suitable for vaginal administration may be presentedas pessaries, tamports, creams, gels, pastes, foams or sprayformulations containing in addition to the active ingredient suchcarriers as are known in the art to be appropriate.

[0553] The compounds may also be entrapped in microcapsules prepared,for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles andnanocapsules) or in macroemulsions. REMINGTON'S PHARMACEUTICAL SCIENCES(A. Osol ed., 16th ed. (1980)).

[0554] In a specific embodiment, the compounds disclosed herein areformulated as liposomes. Liposomes containing a compound of the presentinvention are prepared by methods known in the art. See, e.g. U.S. Pat.Nos. 5,013,556; 4,485,045; 4,544,545; WO 97/38731; Epstein et al., 82PROC. NATL. ACAD. SCI. USA 3688 (1985); and Hwang et al., 77 PROC. NATL.ACAD. SCI. USA 4030 (1980). The compounds of the present invention canalso be administered in the form of liposome delivery systems such assmall unilamellar vesicles, large unilamellar vesicles, andmultilamellar vesicles. Liposomes can be formed from a variety ofphospholipids such as cholesterol, stearylamine or phophatidylcholines.

[0555] Compounds of the present invention may also be delivered by theuse of monoclonal antibodies as individual carriers to which thecompound molecules are coupled. The compounds of the present inventionmay also be coupled with soluble polymers as targetable drug carriers.Such polymers can include polyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamidephenol,polyhydroxyethylaspartamidephenol, or polyethyl-eneoxidepolylysinesubstituted with palmitoyl residue.

[0556] Pharmaceutically Acceptable Preservatives

[0557] The present invention provides stable formulations as well aspreserved solutions and formulations containing a preservative as wellas multi-use preserved formulations suitable for pharmaceutical orveterinary use, comprising at least one compound disclosed herein in apharmaceutically acceptable formulation. Formulations in accordance withthe present invention may optionally contain at least one knownpreservative. Preservatives include, but are not limited to, phenol,m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol,phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol,magnesium chloride (e.g., hexahydrate), alkylparaben (methyl, ethyl,propyl, butyl and the like), benzalkonium chloride, benzethoniumchloride, sodium dehydroacetate and thimerosal, or mixtures thereof inan aqueous diluent. Any suitable concentration or mixture can be used asknown in the art, such as 0.001-5%, or any range or value therein.Non-limiting examples include, no preservative, 0.1-2% m-cresol, 0.1-3%benzyl alcohol, 0.001-0.5% thimerosal, 0.001-2.0% pheno, 0.0005-1.0%alkylparaben(s), and the like.

[0558] Other excipients, e.g., isotonicity agents, buffers,antioxidants, preservative enhancers, can be optionally added to thediluent. An isotonicity agent such as glycerin, is commonly used atknown concentrations. A physiologically tolerated buffer is preferablyadded to provide improved pH control. The formulations can cover a widerange of pHs, such as from about pH 4 to about pH 10, specifically, arange from about pH 5 to about pH 9, and more specifically, a range ofabout 6.0 to about 8.0. In one aspect, the formulations of the presentinvention have pH between about 6.8 and about 7.8. Suitable buffersinclude phosphate buffers, for example, sodium phosphate and phosphatebuffered saline (PBS).

[0559] Other additives, such as a pharmaceutically acceptablesolubilizers like Tween 20 (polyoxyethylene (20) sorbitan monolaurate),Tween 40 (polyoxyethylene (20) sorbitan monopalmitate), Tween 80(polyoxyethylene (20) sorbitan monooleate), Pluronic F68(polyoxyethylene polyoxypropylene block copolymers), and PEG(polyethylene glycol) or non-ionic surfactants such as polysorbate 20 or80 or poloxamer 184 or 188, Pluronic® polyls, other block co-polymers,and chelators such as EDTA and EGTA can optionally be added to thepharmaceutical compositions to reduce aggregation. These additives areparticularly useful if a pump or plastic container is used to administerthe pharmacuetical composition. The presence of pharmaceuticallyacceptable surfactant mitigates the propensity for the composition toaggregate.

[0560] During any of the processes for preparation of the compounds ofthe present invention, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This maybe achieved by means of conventional protecting groups, such as thosedescribed in PROTECTIVE GROUPS IN ORGANIC CHEMISTRY (1973); and GREENEAND WUTS, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS (1991). The protectinggroups may be removed at a convenient subsequent stage using methodsknown from the art.

[0561] Routes of Administration

[0562] The invention further relates to the administration of at leastone compound disclosed herein by the following routes, including, butnot limited to oral, parenteral, subcutaneous, intramuscular,intravenous, intrarticular, intrabronchial, intraabdominal,intracapsular, intracartilaginous, intracavitary, intracelial,intracelebellar, intracerebroventricular, intracolic, intracervical,intragastric, intrahepatic, intramyocardial, intraosteal, intrapelvic,intrapericardiac, intraperitoneal, intrapleural, intraprostatic,intrapulmonary, intrarectal, intrarenal, intraretinal, intraspinal,intrasynovial, intrathoracic, intrauterine, intravesical, bolus,vaginal, rectal, buccal, sublingual, intranasal, iontophoretic means, ortransdermal means.

[0563] Pulmonary/Nasal Administration

[0564] There are a several desirable features of an inhalation devicefor administering a compound of the present invention. For example,delivery by the inhalation device is reliable, reproducible, andaccurate. For pulmonary administration, at least one pharmaceuticalcomposition is delivered in a particle size effective for reaching thelower airways of the lung or sinuses. The inhalation device canoptionally deliver small dry particles, e.g. less than about 10 μm,preferably about 1-5 μm, for good respirability.

[0565] According to the invention, at least one pharmaceuticalcomposition can be delivered by any of a variety of inhalation or nasaldevices known in the art for administration of a therapeutic agent byinhalation. Devices capable of depositing aerosolized formulations inthe sinus cavity or alveoli of a patient include metered dose inhalers,nebulizers, dry powder generators, sprayers, and the like. Other devicessuitable for directing pulmonary or nasal administration are also knownin the art.

[0566] All such devices can be used for the administration of apharmaceutical composition in an aerosol. Such aerosols may compriseeither solutions (both aqueous and non aqueous) or solid particles.Metered dose inhalers like the Ventolin® metered dose inhaler, typicallyuse a propellent gas and require actuation during inspiration. See,e.g., WO 98/35888; WO 94/16970. Dry powder inhalers like Turbuhaler®(Astra), Rotahaler® (Glaxo), Diskus® (Glaxo), Spiros® inhaler (Dura),devices marketed by Inhale Therapeutics, and the Spinhaler® powderinhaler (Fisons), use breath-actuation of a mixed powder. See U.S. Pat.Nos. 5,458,135; 4,668,218; WO 97/25086; WO 94/08552; WO 94/06498; and EP0 237 507, each entirely expressly incorporated herein by reference.Nebulizers like AERx®, Aradigm, the Ultravent® nebulizer (Mallinckrodt),and the Acorn II® nebulizer (Marquest Medical Products), the abovereferences entirely expressly incorporated herein by reference, produceaerosols from solutions, while metered dose inhalers, dry powderinhalers, etc. generate small particle aerosols. These specific examplesof commercially available inhalation devices are intended to be arepresentative of specific devices suitable for the practice of theinvention, and are not intended as limiting the scope of the invention.

[0567] Formulations suitable for nasal administration, wherein thecarrier is a solid, include a coarse powder having a particle size, forexample, in the range of 20 to 500 microns which is administered in themanner in which snuff is administered, i.e., by rapid inhalation throughthe nasal passage from a container of the powder held close up to thenose. Suitable formulations, wherein the carrier is a liquid, foradministration, as for example, a nasal spray or as nasal drops, includeaqueous or oily solutions of the active ingredient.

[0568] A spray comprising a pharmaceutical composition of the presentinvention can be produced by forcing a suspension or solution of acompound disclosed herein through a nozzle under pressure. The nozzlesize and configuration, the applied pressure, and the liquid feed ratecan be chosen to achieve the desired output and particle size. Anelectrospray can be produced, for example, by an electric field inconnection with a capillary or nozzle feed. Advantageously, particles ofat least one compound delivered by a sprayer have a particle size in arange of about less than 1 μm to less than about 20 μm.

[0569] Pharmaceutical compositions of at least one of the compounds ofthe present invention suitable for use with a sprayer typically includea compound disclosed herein in an aqueous solution at a concentration ofabout 0.1 mg to about 100 mg of a compound disclosed herein per ml ofsolution or mg/gm, or any range or value therein, including, but notlimited to, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1,0.2., 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 40, 45, 50, 60, 70, 80, 90 or 100 mg/ml or mg/gm. Thepharmaceutical composition can include agents such as an excipient, abuffer, an isotonicity agent, a preservative, a surfactant, or otherknown agents of pharmaceutical compositions.

[0570] A pharmaceutical composition of the present invention can beadministered by a nebulizer such as a jet nebulizer or an ultrasonicnebulizer. Typically, in a jet nebulizer, a compressed air source isused to create a high-velocity air jet through an orifice. As the gasexpands beyond the nozzle, a low-pressure region is created, which drawsa solution of composition protein through a capillary tube connected toa liquid reservoir. The liquid stream from the capillary tube is shearedinto unstable filaments and droplets as it exits the tube, creating theaerosol. A range of configurations, flow rates, and baffle types can beemployed to achieve the desired performance characteristics from a givenjet nebulizer. In an ultrasonic nebulizer, high-frequency electricalenergy is used to create vibrational, mechanical energy, typicallyemploying a piezoelectric transducer. This energy is transmitted to theformulation of composition protein either directly or through a couplingfluid, creating an aerosol including the composition protein.Advantageously, particles of the pharmaceutical composition delivered bya nebulizer have a particle size range of from about less than 1 μm toless than about 20 μm.

[0571] Pharmaceutical compositions comprising a compound of the presentinvention suitable for use with a nebulizer, either jet or ultrasonic,typically include a concentration of about 0.1 mg to about 100 mg of acompound disclosed herein per ml of solution or mg/gm, or any range orvalue therein including, but not limited to, the individual amountsdisclosed for spray compositions. The pharmaceutical composition caninclude other pharmaceutical agents such as an excipient, a buffer, anisotonicity agent, a preservative, a surfactant, and those known in theart for use in nebulizer administration.

[0572] In a metered dose inhaler (MDI), a propellant, a compound of thepresent invention, and any excipients or other additives are containedin a cannister as a mixture including a liquefied, compressed gas.Actuation of the metering valve releases the mixture as an aerosol,preferably containing a particle size range of from about less than 1 μmto less than about 20 μm.

[0573] The desired aerosol particle size can be obtained by employing aformulation of a compound of the present invention produced by variousmethods known to those of skill in the art including, but not limitedto, jet-milling, spray drying, critical point condensation, and thelike. Suitable metered dose inhalers include those manufactured by 3M orGlaxo and employing a hydrofluorocarbon propellant.

[0574] Pharmaceutical compositions for use with a metered-dose inhalerdevice will generally include a finely divided powder containing acompound disclosed herein as a suspension in a non-aqueous medium, forexample, suspended in a propellant with the aid of a surfactant. Thepropellant can be any conventional material employed for this purposesuch as chlorofluorocarbon, a hydrochlorofluorocarbon, ahydrofluorocarbon, or a hydrocarbon including trichlorofluoromethane,dichlorodifluoromethane, dichlorotetrafluoroethanol and1,1,1,2-tetrafluoroethane, HFA-134a (hydrofluroalkane-134a), HFA-227(hydrofluroalkane-227), or the like. In one embodiment, the propellantis a hydrofluorocarbon. The surfactant can be chosen to stabilize thecompound of the present invention as a suspension in the propellant, toprotect the active agent against chemical degradation, and the like.Suitable surfactants include sorbitan trioleate, soya lecithin, oleicacid, or the like. In some cases solution aerosols are preferred usingsolvents such as ethanol. One of ordinary skill in the art willrecognize that the methods of the present invention can be achieved bypulmonary administration of a compound disclosed herein via devices notdescribed herein.

[0575] For absorption through mucosal surfaces, the compositions andmethods of the present invention for administering a compound disclosedherein include an emulsion comprising a plurality of submicronparticles, a mucoadhesive macromolecule, a bioactive peptide, and anaqueous continuous phase, which promotes absorption through mucosalsurfaces by achieving mucoadhesion of the emulsion particles. See, e.g.,U.S. Pat. No. 5,514,670. Mucous surfaces suitable for application of theemulsions of the present invention can include corneal, conjunctival,buccal, sublingual, nasal, vaginal, pulmonary, abdominal, intestinal,and rectal routes of administration. Pharmaceutical compositions forvaginal or rectal administration such as suppositories, can contain asexcipients, for example, polyalkyleneglycols, vaseline, cocoa butter,and the like. Pharmaceutical composition s for intranasal administrationcan be solid and contain excipients, for example, lactose or can beaqueous or oily solutions of nasal drops. For buccal administration,excipients include sugars, calcium stearate, magnesium stearate,pregelinatined starch, and the like. See, e.g., U.S. Pat. No. 5,849,695.

[0576] In another embodiment, the pharmaceutical compositions of thepresent invention may be administered via transdermal routes using formsof transdermal skin patches well known to those of ordinary skill inthat art. For transdermal administration, a compound of the presentinvention is encapsulated in a delivery device such as a liposome orpolymeric nanoparticles, microparticle, microcapsule, or microspheres(referred to collectively as microparticles unless otherwise stated). Anumber of suitable devices are known, including microparticles made ofsynthetic polymers such as polyhydroxy acids such as polylactic acid,polyglycolic acid and copolymers thereof, polyorthoesters,polyanhydrides, and polyphosphazenes, and natural polymers such ascollagen, polyamino acids, albumin and other proteins, alginate andother polysaccharides, and combinations thereof. See, e.g., U.S. Pat.No. 5,814,599. To be administered in the form of a transdermal deliverysystem, the dosage administration may be, for example, continuous ratherthan intermittent throughout the dosage regimen.

[0577] Formulations suitable for topical administration to the skin maybe presented as ointments, creams, gels and pastes comprising theingredient to be administered in a pharmaceutical acceptable carrier. Apreferred topical delivery system is a transdermal patch comprising acompound of the present invention.

[0578] Topical compositions containing a compound of the presentinvention may be admixed with a variety of carrier materials well knownin the art including alcohols, aloe vera gel, allantoin, glycerine,vitamin A and E oils, mineral oil, PPG2 myristyl propionate and the liketo form, for example, alcoholic solutions, topical cleansers, cleansingcreams, skin gels, skin lotions, and shampoos in cream or gelformulations. Examples of such carriers and methods of formulation maybe found in REMINGTON'S PHARMACEUTICAL SCIENCES (1990). Pharmaceuticalformulations may contain from about 0.005% to about 10% by weight of theactive ingredient. In one embodiment, the pharmaceutical formulationscontain from about 0.01% to 5% by weight of the compound of the presentinvention.

[0579] It can be sometimes desirable to deliver the compounds of thepresent invention to the subject over prolonged periods of time, forexample, for periods of one week to one year from a singleadministration. Certain medical devices may be employed to provide acontinuous intermittent or on demand dosing of a patient. The devicesmay be a pump of diffusion apparatus, or other device containing areservoir of drug and optionally diagnostic or monitoring components toregulate the delivery of the drug. Various slow-release, depot orimplant dosage forms can be utilized. For example, a dosage form cancontain a pharmaceutically acceptable non-toxic salt of compounddisclosed herein that has a low degree of solubility in body fluids, forexample, (a) an acid addition salt with a polybasic acid such asphosphoric acid, sulfuric acid, citric acid, tartaric acid, tannic acid,pamoic acid, alginic acid, polyglutamic acid, naphthalene mono- ordi-sulfonic acids, polygalacturonic acid, and the like; (b) a salt witha polyvalent metal cation such as zinc, calcium, bismuth, barium,magnesium, aluminum, copper, cobalt, nickel, cadmium and the like, orwith an organic cation formed from e.g., N,N′-dibenzyl-ethylenediamineor ethylenediamine; or (c) combinations of (a) and (b) e.g., a zinctannate salt. Additionally, the compounds of the present invention or,preferably, a relatively insoluble salt such as those just described,can be formulated in a gel, for example, an aluminum monostearate gelwith, e.g., sesame oil, suitable for injection. Exemplary salts include,but are not limited to, zinc salts, zinc tannate salts, pamoate salts,and the like. Another type of slow-release depot formulation forinjection would contain the compound or salt dispersed or encapsulatedin a slow degrading, non-toxic, non-antigenic polymer such as apolylactic acid/polyglycolic acid polymer, for example, as described inU.S. Pat. No. 3,773,919. The compounds or relatively insoluble saltsthereof such as those described above can also be formulated incholesterol matrix silastic pellets, particularly for use in animals.Additional slow-release, depot or implant formulations, e.g., gas orliquid liposomes are known in the literature. See, e.g., U.S. Pat. No.5,770,222; SUSTAINED AND CONTROLLED RELEASE DRUG DELIVERY SYSTEMS(1978).

[0580] Other examples include provision of the compounds of the presentinvention to be administered by sustained release delivery systemcontaining a biodegradable composition. The biodegradable compositionmay be composed of a biodegradable, water-coagulable, non-polymericmaterial and a biocompatible, non-toxic organic solvent that is miscibleto dispersible in an aqueous medium. The delivery system may beimplanted at an implant site causing the solvent to dissipate, disperseor leach from the composition into surrounding tissue fluid through aresulting microporous matrix.

[0581] As used herein, the term “implant site” is meant to include asite, in or on which the non-polymeric composition is applied.Implantation or implant site can also include the incorporation of thepharmaceutical composition comprising at least one compound of thepresent invention with a solid device. For example, the pharmaceuticalcomposition is incorporated into a coating on a stent that is implantedinto a subject. Additionally, other solid or biodegradeable materialscan be used as a substrate on which the pharmaceutical composition isapplied. The coated material, comprising the pharmaceutical compositionis then implanted, inserted or is adjacent to the subject or patient.The term “biodegradable” means that the non-polymeric material and/ormatrix of the implant will degrade over time by the action of enzymes,by simple or enzymatically catalyzed hydrolytic action and/or by othersimilar mechanisms in the human body. By “bioerodible,” it is meant thatthe implant matrix will erode or degrade over time due, at least inpart, to contact with substances found in the surrounding tissue fluids,cellular action, and the like. By “bioabsorbable,” it is meant that thenon-polymeric matrix will be broken down and absorbed within the humanbody, for example, by a cell, a tissue, and the like.

[0582] Non-polymeric materials that can be used in the compositiongenerally are those that are biocompatible, substantially insoluble inwater and body fluids, and biodegradable and/or bioerodible. Thenon-polymeric material is capable of being at least partiallysolubilized in a water-soluble organic solvent. The non-polymericmaterials are also capable of coagulating or solidifying to form a solidimplant matrix. The non-polymeric material is combined with a compatibleand suitable organic solvent to form a composition that has the desiredconsistency ranging from watery to viscous to a spreadable putty orpaste.

[0583] Suitable organic solvents are those that are biocompatible,pharmaceutically-acceptable, and will at least partially dissolve thenon-polymeric material. The organic solvent has a solubility in waterranging from miscible to dispersible. Optionally, a pore-forming agentcan be included in the composition to generate additional pores in theimplant matrix. The pore-forming agent can be any organic or inorganic,pharmaceutically-acceptable substance that is substantially soluble inwater or body fluid, and will dissipate from the coagulatingnon-polymeric material and/or the solid matrix of the implant intosurrounding body fluid at the implant site.

[0584] The compounds of the present invention are capable of providing alocal or systemic biological, physiological or therapeutic effect in thebody of an animal. In formulating some pharmaceutical compositionsdescribed herein, the compound is preferably soluble or dispersible inthe non-polymeric composition to form a homogeneous mixture, and uponimplantation, becomes incorporated into the implant matrix. As the solidmatrix degrades over time, the compound is capable of being releasedfrom the matrix into the adjacent tissue fluid, and to the pertinentbody tissue or organ, either adjacent to or distant from the implantsite, preferably at a controlled rate. The release of the compound fromthe matrix may be varied, for example, by the solubility of the compoundin an aqueous medium, the distribution of the compound within thematrix, the size, shape, porosity, and solubility and biodegradabilityof the solid matrix. See e.g. U.S. Pat. No. 5,888,533. The amounts andconcentrations of ingredients in the composition administered to thepatient will generally be effective to accomplish the task intended.

[0585] Compounds of the present invention may be administered bybioactive agent delivery systems containing microparticles suspended ina polymer matrix. The microparticles may be microcapsules, microspheresor nanospheres currently known in the art. The microparticles should becapable of being entrained intact within a polymer that is or becomes agel once inside a biological environment. The microparticles can bebiodegradable or non-biodegradable. Many microencapsulation techniquesused to incorporate a bioactive agent into a microparticle carrier aretaught in the art. See e.g. U.S. Pat. Nos. 4,652,441; 5,100,669;4,438,253; and 5,665,428.

[0586] A preferred polymeric matrix will be biodegradable and exhibitwater solubility at low temperature and will undergo reversible thermalgelation at physiological mammalian body temperatures. The polymericmatrix is capable of releasing the substance entrained within its matrixover time and in a controlled manner. The polymers are graduallydegraded by enzymatic or non-enzymatic hydrolysis in aqueous orphysiological environments. See e.g. U.S. Pat. No. 6,287,588.

[0587] Compounds of the present invention may be administered by a drugdelivery composition comprising microparticles containing at least onechemotherapeutic agent and at least one chemosensitizer suspended in apolymer matrix. The microparticles may be microcapsules, microspheres ornanospheres currently known in the art. The microparticles should bebiodegradable and stable in physiological environments. Themicroparticles also permit diffusion of the chemotherapeutic agent andchemosensitizer from the core through the matrix at a predeterminedrelease rate. Ionic chemotherapeutic agents are suitable for use in thedelivery composition of the invention. Ionic chemosensitizers aresuitable for use in the delivery composition of the invention. The drugdelivery compositions may be delivered to a target site through avariety of known routes of administration. Dosages of thechemotherapeutic agent and chemosensitiThe drug delivery compositionsmay be delivered to a target site through a variety of known routes ofadministration. Dosages of the chemotherapeutic agent andchemosensitizer incorporated in the drug delivery composition willdepend on individual needs, the desired effect and on the chosen routeof administration. See e.g. WO 98/50018.

[0588] Dosage Determinations

[0589] In general, the compounds disclosed herein may be used alone orin concert with other therapeutic agents at appropriate dosages definedby routine testing in order to obtain optimal efficacy while minimizingany potential toxicity. The dosage regimen utilizing a compound of thepresent invention may be selected in accordance with a variety offactors including type, species, age, weight, sex, medical condition ofthe patient; the severity of the condition to be treated; the route ofadministration; the renal and hepatic function of the patient; and theparticular compound employed. A physician or veterinarian of ordinaryskill can readily determine and prescribe the effective amount of thedrug required to prevent, counter, or arrest the progress of thecondition.

[0590] Optimal precision in achieving concentrations of drug within therange that yields maximum efficacy with minimal toxicity may require aregimen based on the kinetics of the compound's availability to one ormore target sites. Distribution, equilibrium, and elimination of a drugmay be considered when determining the optimal concentration for atreatment regimen. The dosages of a compound disclosed herein may beadjusted when combined to achieve desired effects. On the other hand,dosages of these various therapeutic agents may be independentlyoptimized and combined to achieve a synergistic result wherein thepathology is reduced more than it would be if either agent were usedalone.

[0591] In particular, toxicity and therapeutic efficacy of a compounddisclosed herein may be determined by standard pharmaceutical proceduresin cell cultures or experimental animals, e.g., for determining the LD₅₀(the dose lethal to 50% of the population) and the ED₅₀ (the dosetherapeutically effective in 50% of the population). The dose ratiobetween toxic and therapeutic effect is the therapeutic index and it maybe expressed as the ratio LD₅₀/ED₅₀. Compounds exhibiting largetherapeutic indices are preferred except when cytotoxicity of thecompound is the activity or therapeutic outcome that is desired.Although compounds that exhibit toxic side effects may be used, adelivery system can target such compounds to the site of affected tissuein order to minimize potential damage to uninfected cells and, thereby,reduce side effects. Generally, the compounds of the present inventionmay be administered in a manner that maximizes efficacy and minimizestoxicity.

[0592] Data obtained from cell culture assays and animal studies may beused in formulating a range of dosages for use in humans. The dosages ofsuch compounds lies preferably within a range of circulatingconcentrations that include the ED₅₀ with little or no toxicity. Thedosage may vary within this range depending upon the dosage formemployed and the route of administration utilized. For any compound usedin the methods of the invention, the therapeutically effective dose maybe estimated initially from cell culture assays. A dose may beformulated in animal models to achieve a circulating plasmaconcentration range that includes the IC₅₀ (the concentration of thetest compound that achieves a half-maximal inhibition of symptoms) asdetermined in cell culture. Such information may be used to accuratelydetermine useful doses in humans. Levels in plasma may be measured, forexample, by high performance liquid chromatography.

[0593] Moreover, the dosage administration of the pharmaceuticalcompositions of the present invention may be optimized using apharmacokinetic/pharmacodynamic modeling system. For example, one ormore dosage regimens may be chosen and a pharmacokinetic/pharmacodynamicmodel may be used to determine the pharmacokinetic/pharmacodynamicprofile of one or more dosage regimens. Next, one of the dosage regimensfor administration may be selected which achieves the desiredpharmacokinetic/pharmacodynamic response based on the particularpharmacokinetic/pharmacodynamic profile. See WO 00/67776, which isentirely expressly incorporated herein by reference.

[0594] Methods are known in the art for determining effective doses fortherapeutic and prophylactic purposes for the disclosed pharmaceuticalcompositions or the disclosed drug combinations, whether or notformulated in the same composition. For therapeutic purposes, the term“jointly effective amount,” as used herein, means that amount of eachactive compound or pharmaceutical agent, alone or in combination, thatelicits the biological or medicinal response in a tissue system, animalor human that is being sought by a researcher, veterinarian, medicaldoctor or other clinician, which includes alleviation of the symptoms ofthe disease or disorder being treated. For prophylactic purposes (i.e.,inhibiting the onset or progression of a disorder), the term “jointlyeffective amount” refers to that amount of each active compound orpharmaceutical agent, alone or in combination, that inhibits in asubject the onset or progression of a disorder as being sought by aresearcher, veterinarian, medical doctor or other clinician. Thus, thepresent invention provides combinations of two or more therapeuticagents wherein, for example, (a) each therapeutic agent is administeredin an independently therapeutically or prophylactically effectiveamount; (b) at least one therapeutic agent in the combination isadministered in an amount that is sub-therapeutic or subprophylactic ifadministered alone, but is therapeutic or prophylactic when administeredin combination with the second or additional therapeutic agentsaccording to the invention; or (c) both therapeutic agents areadministered in an amount that is subtherapeutic or sub-prophylactic ifadministered alone, but are therapeutic or prophylactic whenadministered together. Combinations of three or more therapeutic agentsare analogously possible. Methods of combination therapy includecoadministration of a single formulation containing all active agents;essentially contemporaneous administration of more than one formulation;and administration of two or more active agents separately formulated.

[0595] Dosages

[0596] More specifically, the pharmaceutical compositions may beadministered in a single daily dose, or the total daily dosage may beadministered in divided doses of two, three, or four times daily. In thecase of oral administration, the daily dosage of the compositions may bevaried over a wide range from about 0.0001 to about 1,000 mg perpatient, per day. The range may more particularly be from about 0.001mg/kg to 10 mg/kg of body weight per day, about 0.1-100 mg, about 1.0-50mg or about 1.0-20 mg per day for adults (at about 60 kg).

[0597] The daily dosage of the pharmaceutical compositions may be variedover a wide range from about 0.01 to about 1000 mg per adult human perday. For oral administration, the pharmaceutical compositions arepreferably provided in the form of tablets containing from about 0.1 mgto about 1000 mg of the compound or 0.1, 0.2, 0.5, 1.0, 2.0, 5.0, 10.0,15.0, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700,800, 900, or 1000 milligrams of the active compound for the symptomaticadjustment of the dosage to the patient to be treated. An effectiveamount of the drug is ordinarily supplied at a dosage level of fromabout 0.1 mg/kg to about 20 mg/kg of body weight per day. In oneembodiment, the range is from about 0.2 mg/kg to about 10 mg/kg of bodyweight per day. In another embodiment, the range is from about 0.5 mg/kgto about 10 mg/kg of body weight per day. The compounds may beadministered on a regimen of about 1 to about 10 times per day.

[0598] In the case of injections, it is usually convenient to give by anintravenous route in an amount of about 0.01-30 mg, about 0.1-20 mg orabout 0.1-10 mg per day to adults (at about 60 kg). In the case of otheranimals, the dose calculated for 60 kg may be administered as well.

[0599] Doses of a compound of the present invention can optionallyinclude 0.0001 to 1,000 mg/kg/administration, or 0.001 to 100.0mg/kg/administration, from 0.01 to 10 mg/kg/administration, from 0.1 to10 mg/kg/administration, including 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7,0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99 and/or 100-500 mg/kg/administration orany range, value or fraction thereof, or to achieve a serumconcentration of 0.1, 0.5, 0.9, 1.0, 1.1, 1.2, 1.5, 1.9, 2.0, 2.5, 2.9,3.0, 3.5, 3.9, 4.0, 4.5, 4.9, 5.0, 5.5, 5.9, 6.0, 6.5, 6.9, 7.0, 7.5,7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11, 11.5, 11.9, 20,12.5, 12.9, 13.0, 13.5, 13.9, 14.0, 14.5, 4.9, 5.0, 5.5., 5.9, 6.0, 6.5,6.9, 7.0, 7.5, 7.9, 8.0, 8.5, 8.9, 9.0, 9.5, 9.9, 10, 10.5, 10.9, 11,11.5, 11.9, 12, 12.5, 12.9, 13.0, 13.5, 13.9, 14, 14.5, 15, 15.5, 15.9,16, 16.5, 16.9, 17, 17.5, 17.9, 18, 18.5, 18.9, 19, 19.5, 19.9, 20,20.5, 20.9, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 96, 100, 200, 300, 400, 500, 600, 700, 800,900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, and/or 5000 μg/mlserum concentration per single or multiple administration or any range,value or fraction thereof.

[0600] As a non-limiting example, treatment of humans or animals can beprovided as a one-time or periodic dosage of a compound of the presentinvention 0.1 to 100 mg/kg such as 0.5, 0.9, 1.0, 1.1, 1.5, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90 or 100 mg/kg, perday, on at least one of day 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, or 40, or alternatively or additionally,at least one of week 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51,or 52, or alternatively or additionally, at least one of 1, 2, 3, 4, 5,6,, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years, or anycombination thereof, using single, infusion or repeated doses.

[0601] Specifically, the pharmaceutical compositions of the presentinvention may be administered at least once a week over the course ofseveral weeks. In one embodiment, the pharmaceutical compositions areadministered at least once a week over several weeks to several months.In another embodiment, the pharmaceutical compositions are administeredonce a week over four to eight weeks. In yet another embodiment, thepharmaceutical compositions are administered once a week over fourweeks.

[0602] More specifically, the pharmaceutical compositions may beadministered at least once a day for about 2 days, at least once a dayfor about 3 days, at least once a day for about 4 days, at least once aday for about 5 days, at least once a day for about 6 days, at leastonce a day for about 7 days, at least once a day for about 8 days, atleast once a day for about 9 days, at least once a day for about 10days, at least once a day for about 11 days, at least once a day forabout 12 days, at least once a day for about 13 days, at least once aday for about 14 days, at least once a day for about 15 days, at leastonce a day for about 16 days, at least once a day for about 17 days, atleast once a day for about 18 days, at least once a day for about 19days, at least once a day for about 20 days, at least once a day forabout 21 days, at least once a day for about 22 days, at least once aday for about 23 days, at least once a day for about 24 days, at leastonce a day for about 25 days, at least once a day for about 26 days, atleast once a day for about 27 days, at least once a day for about 28days, at least once a day for about 29 days, at least once a day forabout 30 days, or at least once a day for about 31 days.

[0603] Alternatively, the pharmaceutical compositions may beadministered about once every day, about once every 2 days, about onceevery 3 days, about once every 4 days, about once every 5 days, aboutonce every 6 days, about once every 7 days, about once every 8 days,about once every 9 days, about once every 10 days, about once every 11days, about once every 12 days, about once every 13 days, about onceevery 14 days, about once every 15 days, about once every 16 days, aboutonce every 17 days, about once every 18 days, about once every 19 days,about once every 20 days, about once every 21 days, about once every 22days, about once every 23 days, about once every 24 days, about onceevery 25 days, about once every 26 days, about once every 27 days, aboutonce every 28 days, about once every 29 days, about once every 30 days,or about once every 31 days.

[0604] The pharmaceutical compositions of the present invention mayalternatively be administered about once every week, about once every 2weeks, about once every 3 weeks, about once every 4 weeks, about onceevery 5 weeks, about once every 6 weeks, about once every 7 weeks, aboutonce every 8 weeks, about once every 9 weeks, about once every 10 weeks,about once every 11 weeks, about once every 12 weeks, about once every13 weeks, about once every 14 weeks, about once every 15 weeks, aboutonce every 16 weeks, about once every 17 weeks, about once every 18weeks, about once every 19 weeks, about once every 20 weeks.

[0605] Alternatively, the pharmaceutical compositions of the presentinvention may be administered about once every month, about once every 2months, about once every 3 months, about once every 4 months, about onceevery 5 months, about once every 6 months, about once every 7 months,about once every 8 months, about once every 9 months, about once every10 months, about once every 11 months, or about once every 12 months.

[0606] Alternatively, the pharmaceutical compositions may beadministered at least once a week for about 2 weeks, at least once aweek for about 3 weeks, at least once a week for about 4 weeks, at leastonce a week for about 5 weeks, at least once a week for about 6 weeks,at least once a week for about 7 weeks, at least once a week for about 8weeks, at least once a week for about 9 weeks, at least once a week forabout 10 weeks, at least once a week for about 11 weeks, at least once aweek for about 12 weeks, at least once a week for about 13 weeks, atleast once a week for about 14 weeks, at least once a week for about 15weeks, at least once a week for about 16 weeks, at least once a week forabout 17 weeks, at least once a week for about 18 weeks, at least once aweek for about 19 weeks, or at least once a week for about 20 weeks.

[0607] Alternatively the pharmaceutical compositions may be administeredat least once a week for about 1 month, at least once a week for about 2months, at least once a week for about 3 months, at least once a weekfor about 4 months, at least once a week for about 5 months, at leastonce a week for about 6 months, at least once a week for about 7 months,at least once a week for about 8 months, at least once a week for about9 months, at least once a week for about 10 months, at least once a weekfor about 11 months, or at least once a week for about 12 months.

[0608] Combination Therapy

[0609] In addition, co-administration or sequential administration ofthe compounds of the present invention and other therapeutic agents maybe desirable, such as chemotherapeutic agents, immunosuppressive agents,cytokines, cytotoxic agents, nucleolytic compounds, radioactiveisotopes, receptors, and pro-drug activating enzymes, which may benaturally occurring or produced by recombinant methods. The combinedadministration includes co-administration, using separate formulationsor a single pharmaceutical formulation, and consecutive administrationin either order, wherein preferably there is a time period while both(or all) active therapeutic agents simultaneously exert their biologicalactivities.

[0610] The compounds of this invention may be administered incombination with at least one selected from the group consisting of anantirheumatic (e.g., methotrexate, auranofin, aurothioglucose,azathioprine, etanercept, gold sodium thiomalate, hydroxychloroquinesulfate, leflunomide, sulfasalzine), a muscle relaxant, a narcotic, anon-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic,a sedative, a local anesthetic, a neuromuscular blocker, an anti-cancer,an antimicrobial (e.g., aminoglycoside, an antifungal, an antiparasitic,an antiviral, a carbapenem, cephalosporin, a flurorquinolone, amacrolide, a penicillin, a sulfonamide, a tetracycline, anotherantimicrobial), an anti-psoriatic, a corticosteriod, an anabolicsteroid, a diabetes-related agent, a mineral, a nutritional, a thyroidagent, a vitamin, a calcium-related hormone, an antidiarrheal, ananti-tussive, an anti-emetic, an anti-ulcer, a laxative, ananticoagulant, an erythropieitin (e.g., epoetin alpha), a filgrastim(e.g., G-CSF, Neupogen), a sargramostim (GM-CSF, Leukine), animmunization, an immunoglobulin, an immunosuppressive (e.g.,basiliximab, cyclosporine, daclizumab), a growth hormone, a hormonereplacement drug, an estrogen receptor modulator, a mydriatic, acycloplegic, an alkylating agent, an anti-metabolite, a mitoticinhibitor, a radiopharmaceutical, an anti-depressant, anti-manic agent,an anti-psychotic, an anxiolytic, a hypnotic, a sympathomimetic, astimulant, donepezil, tacrine, an asthrna medication, a beta agonist, aninhaled steroid, a leukotriene inhibitor, a methylxanthine, a cromolyn,an epinephrine or analog thereof, dornase alpha (Pulmozyme), or acytokine.

[0611] Such anti-cancer or antimicrobial compounds can also includetoxin molecules that are associated, bound, co-formulated,co-administered or sequentially administered, in either order, with atleast one of the compounds of the present invention. The toxin canoptionally act to selectively kill the pathologic cell or tissue. Thepathologic cell can be a cancer or other cell. Such toxins can be, butare not limited to, purified or recombinant toxin or toxin fragmentcomprising at least one functional cytotoxic domain of toxin, e.g.,selected from at least one of ricin, diphtheria toxin, a venom toxin, ora bacterial toxin. The term toxin also includes both endotoxins andexotoxins produced by any naturally occurring, mutant or recombinantbacteria or viruses which may cause any pathological condition in humansand other mammals, including toxin shock, which can result in death.Such toxins may include, but are not limited to, enterotoxigenic E. coliheat-labile enterotoxin (LT), heat-stable enterotoxin (ST), Shigellacytotoxin, Aeromonas enterotoxins, toxic shock syndrome toxin-1(TSST-1), Staphylococcal enterotoxin A (SEA), B (SEB), or C (SEC),Streptococcal enterotoxins and the like. Such bacteria include, but arenot limited to, strains of a species of enterotoxigenic E. coli (ETEC),enterohemorrhagic E. coli (e.g., strains of serotype 0157:H7),Staphylococcus species (e.g., Staphylococcus aureus, Staphylococcuspyogenes), Shigella species (e.g., Shigella dysenteriae, Shigellaflexneri, Shigella boydii, and Shigella sonnei), Salmonella species(e.g., Salmonella typhi, Salmonella cholera-suis, Salmonellaenteritidis), Clostridium species (e.g., Clostridium perfringens,Clostridium dificile, Clostridium botulinum), Camphlobacter species(e.g., Camphlobacter jejuni, Camphlobacter fetus), Heliobacter species,(e.g., Heliobacter pylori), Aeromonas species (e.g., Aeromonas sobria,Aeromonas hydrophila, Acromonas caviae), Plelsomonas shigelloides,Yersina enterocolitica, Vibrios species (e.g., Vibrios cholerae, Vibriosparahemolyticus), Klebsiella species, Pseudomonas aeruginosa, andStreptococci. See, e.g., Stein, ed., INTERNAL MEDICINE, 3rd ed., pp1-13, Little, Brown and Co., Boston, (1990); Evans et al., eds.,Bacterial Infections of Humans: Epidemiology and Control, 2d. Ed., pp239-254, Plenum Medical Book Co., New York (1991); Mandell et al,Principles and Practice of Infectious Diseases, 3d. Ed., ChurchillLivingstone, N.Y. (1990); Berkow et al, eds., The Merck Manual, 16thedition, Merck and Co., Rahway, N.J., 1992; Wood et al, FEMSMicrobiology Immunology, 76:121-134 (1991); Marrack et al, Science,248:705-711 (1990), the contents of which references are incorporatedentirely herein by reference.

[0612] More specifically, the compound of the present invention may beadministered in combination with at least one immunosuppressive agentfor use in, for example, treating or preventing a vascular occlusiveconditions such as transplant vasculopathy. Suitable immunosuppressiveagents include, but are not limited to, CellCept (Roche Labs.), Gengraf(Abbott Labs., Inc.), Micrhogam (Ortho-Clinical), Neoral (Novartis),Orthoclone OKT3 (Ortho-Biotech), Prograf (Fujisawa), Rapamune(Wyeth-Ayerst), Sandimmune (Novartis), Thymoglobulin (SangStat), Zenapax(Roche).

[0613] In one embodiment, the therapeutic agent administeredsimultaneously or sequentially, in either order and at various timeswith a compound of the present invention, comprises a chemotherapeuticagent. A “chemotherapeutic agent” is a compound useful in the treatmentof cancer. Examples of chemotherapeutic agents include, but are notlimited to, alkylating agents such as thiotepa and cyclosphosphamide;alkyl sulfonates such as busulfan, improsulfan and piposulfan;aziridines such as benzodopa, carboquone, meturedopa, and uredopa;ethylenimines and methylamelamines including altretamine,triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide and trimethylolomelamine; nitrogenmustards such as chlorambucil, chlomaphazine, cholophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembiehin, phenesterine, prednimustine,trofosfamide, uracil mustard; nitroureas such as cannustine,chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin,carzinophilin, chromoinycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idambicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such as methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine,5-FU; androgens such as calusterone, dromostanolone propionate,epitiostanol, mepitiostane, testolactone; anti-adrenals such asaminoglutethimide, mitotane, trilostane; folic acid replenisher such asfrolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinicacid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine;demecolcine; diaziquone; elfornithine; elliptinium acetate; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone;mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane;sizofrran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g.,paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) anddoxetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France);chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate;platinum analogs such as cisplatin and carboplatin; vinblastine;platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone;vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin;aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylomithine (DMFO); retinoic acid; esperamicins;capecitabine; and pharmaceutically acceptable salts, acids orderivatives of any of the above. Also included in this definition areanti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens including for example tamoxifen,raloxifene, aromatase inhibiting 4(5)-imidazoles, 4 hydroxytamoxifen,trioxifene, keoxifene, onapristone, and toremifene (Fareston); andanti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide,and goserelin; and pharmaceutically acceptable salts, acids orderivatives of any of the above.

[0614] In another embodiment, the therapeutic agent comprises acytokine. The term “cytokine” is a generic term for proteins released byone cell population which act on another cell as intercellularmediators. Examples of such cytokines are lymphokines, monokines, andtraditional polypeptide hormones. Included among the cytokines aregrowth hormones such as human growth hormone, N-methionyl human growthhormone, and bovine growth hormone; parathyroid hormone; thyroxine;insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such asfollicle stimulating hormone (FSH), thyroid stimulating hormone (TSH),and luteinizing hormone (LH); hepatic growth factor; fibroblast growthfactor; prolactin; placental lactogen; tumor necrosis factor-α and -β;mullerian-inhibiting substance; mouse gonadotropin-associated peptide;inhibin; activin; vascular endothelial growth factor; integrin;thrombopoietin (TPO); nerve growth factors such as NGF-β; plateletgrowth factor; transformning growth factors (TGFs) such as TGF-α andTGF-β; insulin-like growth factor-I and -II; erythropoietin (EPO);osteoinductive factors; interferons such as interferon-α, -β and -γ;colony stimulating factors (CSFs) such as macrophage-CSF (M-CSF);granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (GCSF);interleukins (ILs) such as IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6,IL-7, IL-8, IL-9, IL-11, IL-12, IL-15; a tumor necrosis factor such asTNF-α or TNF-β; and other polypeptide factors including LIF and kitligand (KL). As used herein, the term cytokine includes proteins fromnatural sources or from recombinant cell culture and biologically activeequivalents of the native sequence cytokines.

[0615] In another embodiment, the compounds of the present invention maybe administered in combination with an anti-inflammatory agentincluding, but not limited to, adrenocortical steroids (cortisol,cortisone, fludrocortisone, prednisone, prednisolone,6α-methylprednisolone, triamcinolone, betamethasone, and dexamethasone),non-steroidal agents (salicylic acid derivatives, i.e., aspirin;para-aminophenol derivatives, i.e., acetominophen; indole and indeneacetic acids (indomethacin, sulindac, and etodalac), heteroaryl aceticacids (tolmetin, diclofenac, and ketorolac), arylpropionic acids(ibuprofen and derivatives), anthranilic acids (mefenamic acid, andmeclofenamic acid), enolic acids (piroxicam, tenoxicam, phenylbutazone,and oxyphenthatrazone), nabumetone, gold compounds (auranofin,aurothioglucose, gold sodium thiomalate). Commercially availablenonsteroidal anti-inflammatory drugs include, but are not limited to,Anaprox (Roche Labs.), Arthrotec (Searle), Cataflam (Novartis), Celebrex(Pfizer), , Clinoril (Merck), Dolobid (Merck), Feldene (Pfizer), Indocin(Merck), Lodine (Wyeth-Ayerst), Mobic (Boehringer Ingelheim), Motrin(McNeil Consumer), Naprosyn (Roche Labs.), Orudis (Wyeth-Ayerst),Oruvail (Wyeth-Ayerst), Ponstel (First Horizon), Relafen(GlaxoSmithKline), Tolectin (Ortho-McNeil), Toradol (Roche Labs., Inc.),Vioxx (Merck), Voltaren (Novartis), Advair (GlaxoSmithKline), Flovent(GlaxoSmithKline), Pulmicort (AstranZeneca), and Vanceril (Schering),Asacol (Procter & Gamble), Colazal (Salix), Dipentum (Pharmacia &Upjohn), and Rowasa (Solvay).

[0616] In yet another embodiment, the compounds of the present inventionmay be admistered in combination with an antirheumatic agent.Commercially available antirheumatic agents include, but are not limitedto, Anaprox (Roche Labs.), Arava (Aventic), Arthrotec (Searle),Azulfidine (Pharmacia & Upjohn), Cataflam (Novartis), Celebrex (Pfizer),Celestone (Schering), Cuprimine (Merck), Enbrel (Immunex), Feldene(Pfizer), Gengraf (Abbott), Indocin (Merck), Lodine (Wyeth-Ayerst),Naprosyn (Roche Labs.), Neoral (Novartis), Pediapred (Celltech),Prednisone (Roxanne), Remicade (Centocor), Solu-Medrol (Pharmacia &Upjohn), Triliate (Purdue Frederick), and Voltaren (Novartis).

[0617] Moreover, the compounds of the present invention may be used incombination with any cardiovascular agent including, but not limited to,adrenergic blockers such as Cardura (Pfizer), Dibenzyline (WellSpring),Hytrin (Abbott), Minipress (Pfizer), and Minizide (Pfizer); adrenergicstimulants such as Aldoclor (Merck), Aldomet (Merck), Aldoril (Merck),Catapres (Boehringer Ingelheim), Clorpres (Bertek), and Tenex (Robins);alpha/beta adrenergic blockers such as Coreg (GlaxoSmithKline), andNormodyne (Schering); angiotensin converting enzyme inhibitors such asAccupril (Parke-Davis), Aceon (Solvay), Altace (Monarch), Captopril(Mylan), Enalaprilat (Baxter Anesthesia), Lotensin (Novartis), Mavik(Abbott), Monopril (Bristol-Myers Squibb), Prinivil (Merck), Univasc(Schwarz), Vaotec (Merck), and Zestril (AstraZeneca); angiotenisinconverting enzyme inhibitors such as Lexxel (AstraZeneca), Lotrel(Novartis), Tarka (Abbott), Accuretic (Parke-Davis), Lotensin(Novartis), Prinzide (Merck), Uniretic (Schwarz), Vaeretic (Merck), andZestoretic (AstraZeneca); angiotensin II receptor antagonists such asAtacand (AstraZeneca), Avapro (Briston-Myers Squibb), Cozaar (Merck),Diovan (Novartis), Micardis (Boehringer Ingelheim), and Teveten(Unimed); antiarrhythmics (Groups I-IV), antilipemic agents such as bileacid sequestrants, fibric acid derivatives, HMG-CoA reductaseinhibitors, and nicotinic acid; Beta adrenergic blocking agents; calciumchannel blockers; inotropic agents; vasodilators including coronoaryvasodilators, natriuretic peptides, and peripheral vasodilators; andvasopressors.

[0618] In another aspect of the present invention, the therapeutic agentcomprises a small molecule toxin, including maytansine, calicheamicin,trichothene, and CC 1065. In a specific embodiment, the therapeuticagent may comprise one more calicheamicin molecules. The calicheamicinfamily of antibiotics are capable of producing double-stranded DNAbreaks at sub-picomolar concentrations. Structured analogues ofcalicheamicin are also known. See Hinman et al., 53 CANCER RESEARCH3336-42 (1993); Lode et al., 58 CANCER RESEARCH 2925-28 (1998).

[0619] In yet another aspect of the present invention, the therapeuticagent may comprise one or more enzymatically active toxins and fragmentsthereof. Examples of such toxins include nonbinding active fragments ofdiphtheria toxin, diphtheria A chain, exotoxin A chain (from Pseudomonasaeruginosa), ricin A chain, abrin A chain, modeccin A chain,alpha-sarcin, dianthin proteins, Phytolaca americana proteins (PAPI,PAPAII, and PAP-S), momordica charantia inhibitor, curcin, crotinsapaonaria officinalis inhibitor, gelonin, mitogellin, restrictoein,phenomvcin, enomycin and the tricothecenes. See, e.g., WO 93/21232.

[0620] The present invention further contemplates therapeutic agentsthat have nucleolytic activity such as a ribonuclease and adeoxyribonuclease. In addition, a variety of radioactive isotopes areavailable for the production of radioconjugated binding partners.Examples include Y⁹⁰, At²²², Ret⁸⁶, Re¹⁸⁶, Sm¹⁵³, Bi²¹², P³² andradioactive isotopes of Lu.

[0621] In yet another aspect of the present invention, the at least onecompound may be conjugated to a receptor, such as streptavidin, forutilization in tumor pretargeting. Briefly, the compound-receptorconjugate is administered to the patient and unbound conjugate isremoved from circulation with a clearing agent. A ligand, such asbiotin, which is conjugated to a cytotoxic agent is then administered.

[0622] Timing of Administration

[0623] In several embodiments of the present invention, a compounddescribed herein is administered before or after administration of asecond therapeutic agent. The administration of a compound may occuranytime from several minutes to several hours before the administrationof the second therapeutic agent. The compound may alternatively beadministered anytime from several hours to several days, possiblyseveral weeks, and up to several months before the second therapeuticagent.

[0624] More specifically, a compound of the present invention may beadministered at least about 1 minute, at least about minutes, at leastabout minutes, at least about minutes, at least about minutes, at leastabout 2 minutes, at least about 3 minutes, at least about 4 minutes, atleast about 5 minutes, at least about 6 minutes, at least about 7minutes, at least about 8 minutes, at least about 9 minutes, at leastabout 10 minutes, at least about 11 minutes, at least about 12 minutes,at least about 13 minutes, at least about 14 minutes, at least about 15minutes, at least about 16 minutes, at least about 17 minutes, at leastabout 18 minutes, at least about 19 minutes, at least about 20 minutes,at least about 21 minutes, at least about 22 minutes, at least about 23minutes, at least about 24 minutes, at least about 25 minutes, at leastabout 26 minutes, at least about 27 minutes, at least about 28 minutes,at least about 29 minutes, at least about 30 minutes, at least about 31minutes, at least about 32 minutes, at least about 33 minutes, at leastabout 34 minutes, at least about 35 minutes, at least about 36 minutes,at least about 37 minutes, at least about 38 minutes, at least about 39minutes, at least about 40 minutes, at least about 41 minutes, at leastabout 42 minutes, at least about 43 minutes, at least about 44 minutes,at least about 45 minutes, at least about 46 minutes, at least about 47minutes, at least about 48 minutes, at least about 49 minutes, at leastabout 50 minutes, at least about 51 minutes, at least about 52 minutes,at least about 53 minutes, at least about 54 minutes, at least about 55minutes, at least about 56 minutes, at least about 57 minutes, at leastabout 58 minutes, at least about 59 minutes, or at least about 60minutes before or after the second therapeutic agent. Furthermore, acompound of the present invention may be administered at least about 1hour, at least about 2 hours, at least about 3 hours, at least about 4hours, at least about 5 hours, at least about 6 hours, at least about 7hours, at least about 8 hours, at least about 9 hours, at least about 10hours, at least about 11 hours, at least about 12 hours, at least about13 hours, at least about 14 hours, at least about 15 hours, at leastabout 16 hours, at least about 17 hours, at least about 18 hours, atleast about 19 hours, at least about 20 hours, at least about 21 hours,at least about 22 hours, at least about 23 hours, or at least about 24hours before or after the second therapeutic agent.

[0625] Moreover, a compound of the present invention may be administeredat least about 1 day, at least about 2 days, at least about 3 days, atleast about 4 days, at least about 5 days, at least about 6 days, atleast about 7 days, at least about 8 days, at least about 9 days, atleast about 10 days, at least about 11 days, at least about 12 days, atleast about 13 days, at least about 14 days, at least about 15 days, atleast about 16 days, at least about 17 days, at least about 18 days, atleast about 19 days, at least about 20 days, at least about 21 days, atleast about 22 days, at least about 23 days, at least about 24 days, atleast about 25 days, at least about 26 days, at least about 27 days, atleast about 28 days, at least about 29 days, at least about 30 days orat least about 31 days before or after the administration of the secondtherapeutic agent.

[0626] In yet another aspect of the present invention, a compound of thepresent invention may be administered at least about 1 week, at leastabout 2 weeks, at least about 3 weeks, at least about 4 weeks, at leastabout 5 weeks, at least about 6 weeks, at least about 7 weeks, at leastabout 8 weeks, at least about 9 weeks, at least about 10 weeks, at leastabout 11 weeks, at least about 12 weeks, at least about 13 weeks, atleast about 14 weeks, at least about 15 weeks, at least about 16 weeks,at least about 17 weeks, at least about 18 weeks, at least about 19weeks, or at least about 20 weeks before or after the second therapeuticagent.

[0627] In a further aspect of the present invention, a compound of thepresent invention may be administered at least about one month, at leastabout two months, at least about three months, at least about fourmonths, at least about five months, at least about six months, at leastabout seven months, at least about eight months, at least about ninemonths, at least about ten months, at least about eleven months, or atleast about twelve months before or after the second therapeutic agent.

[0628] For convenience, the meaning of certain terms and phrasesemployed in the specification, examples, and appended claims areprovided below.

[0629] Definitions

[0630] As used herein, the term “compound” includes both the singularand the plural, and includes any single entity or combined entities thathave at least the activity disclosed herein and combinations, fragments,analogs or derivatives of such entities. Such entities include, but arenot limited to, chemical elements, molecules, compounds, mixtures,emulsions, chemotherapeutic agents, pharmacological agents, hormones,antibodies, growth factors, cellular factors, nucleic acids, proteins,peptides, peptidomimetics, nucleotides, carbohydrates, and combinations,fragments, analogs or derivatives of such entities.

[0631] The term “phenylamine” refers to a primary or secondarybenzeneamine, more commonly known as an aniline. The amino group on theaniline can be substituted with hydrogen, alkyl (C₁-C₁₂, straight chainor branched), cycloalkyl (C₃-C₁₀), or aryl substituted aryl groups. Thephenyl ring of this aniline derivative can be optionally substitutedwith one or more functional groups, or a combination of functionalgroups such as alkyl, alkenyl, alkynyl, phenyl, benzyl, halo, cyano,nitro, hydroxy, thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio,arylthio, amino, alkyl amino, aryl amino, acyl, carboxyl, amido,sulfonamido, sulfonyl, sulfate, sulfonic acid, morpholino, piperazinyl,pyridyl, thienyl, furanyl, pyrroyl, pyrazoyl, phosphate, phosphonicacid, or phosphonate. If applicable, these groups can be represented inprotected or unprotected forms used in standard organic synthesis.

[0632] The term “naphthylamine” refers to a primary or secondary a- orβ-naphthylamine. The ring substructure in the naphthylamine can beoptionally substituted with one or a combination of functional groupssuch as alkyl, alkenyl, alkynyl, phenyl, benzyl, halo, cyano, nitro,hydroxy, thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio, arylthio,amino, alkyl amino, aryl amino, acyl, carboxyl, amido, sulfonamido,sulfonyl, sulfate, sulfonic acid, morpholino, thiomorpholino,piperazinyl, pyridyl, thienyl, furanyl, pyrroyl, pyrazoyl, phosphate,phosphonic acid, phosphonate and the like. These groups can berepresented in protected or unprotected forms used in standard organicsynthesis.

[0633] The term “naphthylalkyl amine” refers to a primary or secondaryα- and β-naphthylalkyl amine (for example, 2-α-naphthylethyl amine). Theterm “benzalkyl amine” refers to a primary or secondary benzylalkylamine (for example, phenylethyl amine). These aryl alkyl substructuresor compounds can be optically active or optically inactive. The aryl(ring) substructures of the naphthylalkyl and benzalkyl amines can beoptionally subsituted with one or a combination of functional groups,such as alkyl, alkenyl, alkynyl, phenyl, benzyl, halo, cyano, nitro,hydroxy, thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio, arylthio,amino, alkyl amino, aryl amino, acyl, carbolyl, amido, sulfonamido,sulfonyl, sulfate, sulfonic acid, morpholino, piperazinyl, pyridyl,thienyl, ftiranyl, pyrroyl, pyrazoyl, phosphate, phosphonic acid,phosphonate and the like. If applicable these groups can be representedin protected or unprotected forms used in standard organic synthesis.

[0634] The term “quinolinyl amine” refers to primary or secondaryquinolyl amines. These amines can be in optically active or inactiveforms. The aryl (ring) substructure of the quinolyl amine can be beoptionally substituted with one a combination of functional groups suchas alkyl, alkenyl, alkynyl, phenyl, benzyl, halo, cyano, nitro, hydroxy,thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio, arylthio, amino, alkylamino, aryl amino, acyl, carboxyl, amido, sulfonamido, sulfonyl,sulfate, sulfonic acid, morpholino, thiomorpholino, piperazinyl,pyridyl, thienyl, furanyl, pyrroyl, pyrazoyl, phosphate, phosphonicacid, phosphonate and the like. These groups can be represented inprotected or unprotected forms used in standard organic synthesis.

[0635] The term “heteroaryl amines” refers to pyrroles, pyrazoles,imidazoles, and indoles. The aryl (ring) substructure of the heteroarylamine can be optionally substituted with one or a combination offunctional groups such as alkyl, alkenyl, alkynyl, phenyl, benzyl, halo,cyano, nitro, hydroxy, thioxy, alkoxy, aryloxy, haloalkyloxy, alkylthio,arylthio, amino, alkyl amino, aryl amino, acyl, carboxyl, amido,sulfonamido, sulfonyl, sulfate, sulfonic acid, morpholino,thiomorpholino, piperazinyl, phosphate, phosphonic acid, or phosphonate.These groups can be represented in protected or unprotected forms usedin standard organic synthesis.

[0636] The term “glycated protein,” as used herein, includes proteinslinked to glucose, either enzymatically or non-enzymatically, primarilyby condensation of free epsilon-amino groups in the protein withglucose, forming Amadori adducts. Furthermore, glycated protein, as usedherein, includes not only proteins containing these initial glycationproducts, but also glycation products resulting from further reactionssuch as rearrangements, dehydration, and condensations that formirreversible advanced glycation end products (AGE).

[0637] The term “polynucleotide” refers generally to polymeric forms ofnucleotides of any length, either ribonucleotides or deoxynucleotides.Thus, this term includes, but is not limited to, single-stranded,double-stranded, or multi-stranded DNA or RNA. Polynucleotides mayfurther comprise genomic DNA, cDNA, or DNA-RNA hybrids. Moreover, thepolynucleotides of the present invention may be synthetically produced.

[0638] Polynucleotides may comprise chemically modified, biochemicallymodified, or derivatized nucleotides. For example, a polynucleotide maycomprise, in part, modified nucleotides such as methylated nucleotidesor nucleotide analogs. In other embodiments, polynucleotides maycomprise sugars, caps, nucleotide branches, and linking groups such asfluororibose and thioate. In addition, the sequence of nucleotides maybe interrupted by non-nucleotide components. Furthermore, apolynucleotide may be modified after polymerization to facilitate itsattachment to other polynucleotides, proteins, metal ions, labelingcomponents, or a solid support.

[0639] The backbone of the polynucleotide may comprise modified orsubstituted sugar and/or phosphate groups. Alternatively, the backboneof the polynucleotide may comprise a polymer of synthetic subunits suchas phosphoramidites and thus may be an oligodeoxynucleosidephosphoramidate or a mixed phosphoramidate-phosphodiester oligomer. SeePeyrottes et al., NUCL. ACIDS RES. (1996) 24:1841-1848, and Chaturvediet al., NUCL. ACIDS RES. (1996) 24:2318-2323.

[0640] The term “homology”, as used herein, refers to a degree ofcomplementarity. There may be partial homology or complete homology(i.e., identity). A partially complementary sequence is one that atleast partially inhibits an identical sequence from hybridizing to atarget polynucleotide; it is referred to using the functional term“substantially homologous” The inhibition of hybridization of thecompletely complementary sequence to the target sequence may be examinedusing a hybridization assay (Southern or Northern blot, solutionhybridization and the like) under conditions of low stringency. Asubstantially homologous sequence or probe will compete for and inhibitthe binding (i.e., the hybridization) of a completely homologoussequence or probe to the target sequence under conditions of lowstringency. This is not to say that conditions of low stringency aresuch that non-specific binding is permitted; low stringency conditionsrequire that the binding of two sequences to one another be a specific(i.e., selective) interaction. The absence of non-specific binding maybe tested by the use of a second target sequence which lacks even apartial degree of complementarity (e.g., less than about 30% identity);in the absence of non-specific binding, the probe will not hybridize tothe second non-complementary target sequence.

[0641] The term “gene” refers to a polynucleotide sequence thatcomprises coding sequences necessary for the production of a polypeptideor precursor, and may also include expression control sequences or othercontrol or regulatory sequences. The polypeptide can be encoded by afull length coding sequence or by any portion of the coding sequence.The gene may be derived in whole or in part from any source known tothose of ordinary skill in the art including a plant, a fungus, ananimal, a bacterial genome or episome, eukaryotic, nuclear or plasmidDNA, cDNA, viral DNA, or chemically synthesized DNA. A gene mayconstitute an uninterrupted coding sequence or it may include one ormore introns, bound by the appropriate splice junctions. Moreover, agene may contain one or more modifications in either the coding or theuntranslated regions that could affect certain properties of thepolynucleotide or polypeptide, such as the biological activity or thechemical structure of the expression product, the rate of expression, orthe manner of expression control. Such modifications include, but arenot limited to, mutations, insertions, deletions, and substitutions ofone or more nucleotides. In this regard, such modified genes may bereferred to as variants of the native gene.

[0642] “Gene expression” refers to the process by which a polynucleotidesequence undergoes successful transcription and translation such thatdetectable levels of the nucleotide sequence are expressed as proteinsor the polynucleotide sequence undergoes transcription, if RNA is copiedfrom DNA, or replication if DNA is copied from DNA, such that theresulting nucleotide copies are detectable.

[0643] The term “gene expression profile” refers to a group of genesrepresenting a particular cell or tissue type (e.g., neuron, coronaryartery endothelium, or disease tissue) in any activation state. In oneaspect, a gene expression profile is generated from cells exposed to acompound of the present invention. This profile may be compared to agene expression profile generated from the same type of cell or tissuetype prior to treatment with a compound of the present invention.Furthermore, a series of gene expression profiles may be generated fromcells or tissues treated with a compound of the present invention,specifically, at different doses or a time-course to assess the effectsof the compound. A gene expression profile is also known as a geneexpression signature.

[0644] The term “differential expression” refers to both quantitative aswell as qualitative differences in the temporal and tissue expressionpatterns of a gene. For example, a differentially expressed gene mayhave its expression activated or completely inactivated in normal versusdisease conditions. Such a qualitatively regulated gene may exhibit anexpression pattern within a given tissue or cell type that is detectablein either control or disease conditions, but is not detectable in both.“Differentially expressed polynucleotide,” as used herein, refers to apolynucleotide sequence that uniquely identifies a differentiallyexpressed gene so that detection of the differentially expressedpolynucleotide in a sample is correlated with the presence of adifferentially expressed gene in a sample.

[0645] Similarly, a differentially expressed protein may have itsexpression activated or completely inactivated in normal versus diseaseconditions. Such a qualitatively regulated protein may exhibit anexpression pattern within a given tissue or cell type that is detectablein either control or disease conditions, but is not detectable in both.A “differentially expressed protein,” as used herein, refers to an aminoacid sequence that uniquely identifies a differentially expressedprotein so that detection of the differentially expressed protein in asample is correlated with the presence of a differentially expressedprotein in a sample.

[0646] “Cell type,” as used herein, refers to a cell from a given source(e.g., tissue or organ), a cell in a given state of differentiation, ora cell associated with a given pathology or genetic makeup.

[0647] The term “polypeptide” refers to a polymeric form of amino acidsof any length, which may include translated, untranslated, chemicallymodified, biochemically modified, and derivatized amino acids. Apolypeptide may be naturally occurring, recombinant, or synthetic, orany combination of these. Moreover, the term “polypeptide,” as usedherein, refers to proteins, polypeptides, and peptides of any size,structure, or function. For example, a polypeptide may comprise a stringof amino acids held together by peptide bonds. A polypeptide mayalternatively comprise a long chain of amino acids held together bypeptide bonds. Moreover, a polypeptide may also comprise a fragment of anaturally occurring protein or peptide. A polypeptide may be a singlemolecule or may be a multi-molecular complex. In addition, suchpolypeptides may have modified peptide backbones as well.

[0648] The term “polypeptide” further comprises immunologically taggedproteins and fusion proteins, including, but not limited to, fusionproteins with a heterologous amino acid sequence, fusion proteins withheterologous and homologous leader sequences, and fusion proteins withor without N-terminal methionine residues.

[0649] The term “protein expression” refers to the process by which apolynucleotide sequence undergoes successful transcription andtranslation such that detectable levels of the amino acid sequence orprotein are expressed.

[0650] The term “protein expression profile” refers to a group ofproteins representing a particular cell or tissue type (e.g., neuron,coronary artery endothelium, or disease tissue). In one aspect, aprotein expression profile is generated from cells or tissues exposed toa compound of the present invention. This profile may be compared to aprotein expression profile generated from the same type of cell ortissue prior to treatment with a compound of the present invention.Furthermore, a series of protein expression profiles may be generatedfrom cells or tissues treated with a compound of the present invention,specifically, at different doses or a time-course to assess the effectsof the compound. A protein expression profile is also known as a“protein expression signature.”

[0651] As used herein, a “biomolecule” includes polynucleotides andpolypeptides. Moreover, a “biomolecular sequence,” as used herein, is aterm that refers to all or a portion of a polynucleotide sequence. Abiomolecular sequence may also refer to all or a portion of apolypeptide sequence. In the context of biomolecule, for example,perlecan, the term “functional equivalent” refers to a protein orpolynucleotide molecule that possesses functional or structuralcharacteristics that are substantially similar to all or part of thenative perlecan protein or native perlecan-encoding polynucleotides. Afunctional equivalent of a native perlecan protein may containmodifications depending on the necessity of such modifications for aspecific structure or the performance of a specific function. The term“functional equivalent” is intended to include the “fragments,”“mutants,” “derivatives,” “alleles,” “hybrids,” “variants,” “analogs,”or “chemical derivatives” of native perlecan.

[0652] A “host cell,” as used herein, refers to a microorganism, aprokaryotic cell, a eukaryotic cell or cell line cultured as aunicellular entity that may be, or has been, used as a recipient for arecombinant vector or other transfer of polynucleotides, and includesthe progeny of the original cell that has been transfected. It isunderstood that the progeny of a single cell may not necessarily becompletely identical in morphology or in genomic or total DNA complementas the original parent due to natural, accidental, or deliberatemutation.

[0653] In the context of immunoglobulins, the term “functionalequivalent” refers to immunoglobulin molecules that exhibitimmunological binding properties that are substantially similar to theparent immunoglobulin. As used herein, the term “immunological bindingproperties” refers to non-covalent interactions of the type which occurbetween an immunoglobulin molecule and an antigen for which theimmunoglobulin is specific. Indeed, a functional equivalent of amonoclonal antibody immunoglobulin, for example, may inhibit the bindingof the parent monoclonal antibody to its antigen. A functionalequivalent may comprise F(ab′)₂ fragments, F(ab) molecules, Fvfragments, single chain fragment variable displayed on phage (scFv),single domain antibodies, chimeric antibodies, or the like so long asthe immunoglobulin exhibits the characteristics of the parentimmunoglobulin.

[0654] As used herein, the term “isolated” refers to a polynucleotide, apolypeptide, an antibody, or a host cell that is in an environmentdifferent from that in which the polynucleotide, the polypeptide, theantibody, or the host cell naturally occurs. An isolated polynucleotide,polypeptide, antibody, or host cell is generally substantially purified.

[0655] As used herein, the term “substantially purified” refers to acompound that is removed from its natural environment and is at leastabout 60% to 99.9% free from other components, or is at least about 60%free, at least about 65% free, at least about 70% free, at least about75% free, at least about 80% free, at least about 83% free, at leastabout 85% free, at least about 88% free, at least about 90% free, atleast about 91% free, at least about 92% free, at least about 93% free,at least about 94% free, at least about 95% free, at least about 96%free, at least about 97% free, at least about 98% free, at least about99% free, at least about 99.9% free, or at least about 99.99% free fromother components with which it is naturally associated. For example, acomposition containing A is “substantially free of” B when at leastabout 85% by weight of the total A+B in the composition is A.Alternatively, A comprises at least about 90% by weight of the total ofA+B in the composition, further still, at least about 95% or even 99% byweight.

[0656] “Diagnosis,” as used herein, generally includes a determinationof a subject's susceptibility to a disease or disorder, a determinationas to whether a subject is presently affected by a disease or disorder,a prognosis of a subject affected by a disease or disorder (e.g.,identification of pre-metastatic or metastatic cancerous states, stagesof cancer, or responsiveness of cancer to therapy), and therametrics(e.g., monitoring a subject's condition to provide information as to theeffect or efficacy of therapy).

[0657] The term “biological sample” encompasses a variety of sampletypes obtained from or originating from an organism which may be used indiagnostic, monitoring, or other assays. The term encompasses blood,serum, plasma, cells, proteins, carbohydrates, nucleic acids, urine,nasal secretions, mucosal secretions, cellular fluid, cellular exudateand other liquid samples of biological origin, solid tissue samples suchas a biopsy specimen, or tissue cultures or cells derived therefrom andthe progeny thereof. The term specifically encompasses a clinicalsample, and further includes cells in cell culture, cell supernatants,cell lysates, amniotic fluid, biological fluids, and tissue samples. Theterm also encompasses samples that have been manipulated in any wayafter procurement such as treatment with reagents, solubilization, orenrichment for certain components. The biological sample can be derivedfrom the organism directly or can be collected from the environment.

[0658] The terms “individual,” “subject,” “host,” and “patient” refer toany subject for whom diagnosis, treatment, or therapy is desired. In oneembodiment, the individual, subject, host, or patient is a human. Othersubjects may include, but are not limited to, animals including but notlimited to, cattle, sheep, horses, dogs, cats, guinea pigs, rabbits,rats, primates, opossums and mice. Other subjects include species ofbacteria, phages, cell cultures, viruses, plants and other eucaryotes,prokaryotes and unclassified organisms.

[0659] The terms “treatment,” “treating,” “treat,” and the like are usedherein to refer generally to obtaining a desired pharmacological and/orphysiological effect. The effect may be prophylactic in terms ofcompletely or partially preventing a disease or symptom thereof and/ormay be therapeutic in terms of a partial or complete stabilization orcure for a disease and/or adverse effect attributable to the disease.“Treatment” as used herein covers any treatment of a disease in asubject, particularly a human, and includes: (a) preventing the diseaseor symptom from occurring in a subject which may be predisposed to thedisease or symptom, but has not yet been diagnosed as having it; (b)inhibiting the disease symptom, i.e., arresting its development; or (c)relieving the disease symptom, i.e., causing regression of the diseaseor symptom.

[0660] The expression “therapeutically effective amount” refers to anamount of, for example, a compound disclosed herein, that is effectivefor preventing, ameliorating, treating or delaying the onset of adisease or condition.

[0661] A “prophylactically effective amount” refers to an amount of, forexample, a compound disclosed herein that is effective for preventing adisease or condition.

[0662] A “liposome” is a small vesicle composed of various types oflipids, phospholipids and/or surfactant, which is useful for delivery ofa drug to a subject, such as a mammal or other animal. The compounds ofthe present invention may be delivered by a liposome. The components ofthe liposome are commonly arranged in a bilayer formation, similar tothe lipid arrangement of biological membranes. Liposome formulations,loading of liposomes and administration and delivery of liposomes areknown in the art.

[0663] “Hybridization,” broadly defined, refers to any process by whicha polynucleotide sequence binds to a complementary sequence through basepairing. Hybridization conditions can be defined by, for example, theconcentrations of salt or formamide in the prehybridization andhybridization solutions, or by the hybridization temperature, and arewell known in the art. Hybridization can occur under conditions ofvarious stringency. Hybridization may also refer to the binding of aprotein-capture agent to a target protein under certain conditions, suchas normal physiological conditions.

[0664] As understood herein, the term “activation” refers to anyalteration of a signaling pathway or biological response including, forexample, increases above basal levels, restoration to basal levels froman inhibited state, and stimulation of the pathway above basal levels.

[0665] The term “biological activity” refers to the biological behaviorand effects of a protein or peptide. The biological activity of aprotein may be affected at the cellular level and the molecular level.For example, an antisense oligonucleotide may prevent translation of aparticular mRNA, thereby inhibiting the biological activity of theprotein encoded by the mRNA. In addition, an antibody may bind to aparticular protein and inhibit that protein's biological activity.

[0666] The term “oligonucleotide” as used herein refers to apolynucleotide sequence comprising, for example, from about 4nucleotides (nt) to about 1000 nt. Oligonucleotides for use in thepresent invention are preferably from about 15 nt to about 150 nt, morepreferably from about 150 nt to about 1000 nt in length. Theoligonucleotide may be a naturally occurring oligonucleotide or asynthetic oligonucleotide. Oligonucleotides may be prepared by thephosphoramidite method (Beaucage and Carruthers, TETRAHEDRON LETT.(1981) 22:1859-1862), or by the triester method (Matteucci et al., J.AM. CHEM. SOC. (1981) 103:3185), or by other chemical methods known inthe art.

[0667] The term “microarray” refers generally to the type of genes orproteins represented on a microarray by oligonucleotides (polynucleotidesequences) or protein-binding agents, and where the type of genes orproteins represented on the microarray is dependent on the intendedpurpose of the microarray (e.g., to monitor expression of human genes orproteins). The oligonucleotides or protein-binding agents on a givenmicroarray may correspond to the same type, category, or group of genesor proteins. Genes or proteins may be considered to be of the same typeif they share some common characteristics such as species of origin(e.g., human, mouse, rat); disease state (e.g., cancer); function (e.g.,protein kinases, tumor suppressors); same biological process (e.g.,apoptosis, signal transduction, cell cycle regulation, proliferation,differentiation). For example, one microarray type may be a “cancermicroarray” in which each of the microarray oligonucleotides orprotein-binding agents correspond to a gene or protein associated with acancer. An “epithelial microarray” may be a microarray ofoligonucleotides or protein-binding agents corresponding to uniqueepithelial genes or proteins. Similarly, a “cell cycle microarray” maybe an microarray type in which the oligonucleotides or protein-bindingagents correspond to unique genes or proteins associated with the cellcycle.

[0668] The term “detectable”, one in sense, refers to a polynucleotideexpression pattern which is detectable via the standard techniques ofpolymerase chain reaction (PCR), reverse transcriptase (RT) -PCR(RT-PCR), differential display, and Northern analyses, which are wellknown to those of skill in the art. Similarly, polypeptide expressionpatterns may be “detected” via standard techniques includingimmunoassays such as Western blots. In general, the term “detectable isused when a result of an action, such as addition of a compound in anassay step, is observable, particularly by physical means, such as acolor change.

[0669] A “target gene” refers to a polynucleotide, often derived from abiological sample, to which an oligonucleotide probe is designed tospecifically hybridize. It is either the presence or absence of thetarget polynucleotide that is to be detected, or the amount of thetarget polynucleotide that is to be quantified. The targetpolynucleotide has a sequence that is complementary to thepolynucleotide sequence of the corresponding probe directed to thetarget. The target polynucleotide may also refer to the specificsubsequence of a larger polynucleotide to which the probe is directed orto the overall sequence (e.g., gene or mRNA) whose expression level itis desired to detect.

[0670] A “target protein” refers to an polypeptide, often derived from abiological sample, to which a protein-capture agent specificallyhybridizes or binds. It is either the presence or absence of the targetprotein that is to be detected, or the amount of the target protein thatis to be quantified. The target protein has a structure that isrecognized by the corresponding protein-capture agent directed to thetarget. The target protein or amino acid may also refer to the specificsubstructure of a larger protein to which the protein-capture agent isdirected or to the overall structure (e.g., gene or mRNA) whoseexpression level it is desired to detect.

[0671] The term “complementary” refers to the topological compatibilityor matching together of the interacting surfaces of a probe molecule andits target. The target and its probe can be described as complementary,and furthermore, the contact surface characteristics are complementaryto each other. Hybridization or base pairing between nucleotides ornucleic acids, such as, for example, between the two strands of adouble-stranded DNA molecule or between an oligonucleotide probe and atarget are complementary.

[0672] The term “background” refers to non-specific binding or otherinteractions between, for example, polynucleotides, polypeptides, smallmolecules and polypeptides, or small molecules and polynucleotides.“Background” may also refer to the non-specific binding or otherinteractions in the context of assays including immunoassays.

[0673] In the context of microarrays, the term “background” refers tohybridization signals resulting from non-specific binding, or otherinteractions, between the labeled target polynucleotides and componentsof the oligonucleotide microarray (e.g., the oligonucleotide probes,control probes, the microarray support) or between target proteins andthe protein-binding agents of a protein microarray. Background signalsmay also be produced by intrinsic fluorescence of the microarraycomponents themselves. A single background signal may be calculated forthe entire microarray, or a different background signal may becalculated for each target polynucleotide or target protein. Thebackground may be calculated as the average hybridization signalintensity, or where a different background signal is calculated for eachtarget gene or target protein. Alternatively, background may becalculated as the average hybridization signal intensity produced byhybridization to probes that are not complementary to any sequence foundin the sample (e.g., probes directed to polynucleotides of the oppositesense or to genes not found in the sample such as bacterial genes wherethe sample is mammalian polynucleotides). The background can also becalculated as the average signal intensity produced by regions of themicroarray which lack any probes or protein-binding agents at all.

[0674] A “small molecule” comprises a compound or molecular complex,either synthetic, naturally derived, or partially synthetic, composed ofcarbon, hydrogen, oxygen, and nitrogen, which may also contain otherelements, and which may have a molecular weight of less than about 100to about 15,000 Daltons, or less than about 15,000, less than about14,000, less than about 13,000, less than about 12,000, less than about11,000, less than about 10,000, less than about 9,000, less than about8,000, less than about 7,000, less than about 6,000, less than about5,000, less than about 4,000, less than about 3,000, less than about2,000, less than about 1,000, less than about 900, less than about 800,less than about 700, less than about 600, less than about 500, less thanabout 400, less than about 300, less than about 200, or less than about100.

[0675] The term “fusion protein” refers to a protein composed of two ormore polypeptides that, although typically not joined in their nativestate, are joined by their respective amino and carboxyl termini througha peptide linkage to form a single continuous polypeptide. It isunderstood that the two or more polypeptide components can either bedirectly joined or indirectly joined through a peptide linker/spacer.

[0676] The term “normal physiological conditions” means conditions thatare typical inside a living organism or a cell. Although some organs ororganisms provide extreme conditions, the intra-organismal andintra-cellular environment normally varies around pH 7 (i.e., from pH6.5 to pH 7.5), contains water as the predominant solvent, and exists ata temperature above 0° C. and below 50° C. The concentration of varioussalts depends on the organ, organism, cell, or cellular compartment usedas a reference.

[0677] The term “cluster” refers to a group of clones or biomolecularsequences related to one another by sequence homology. In one example,clusters are formed based upon a specified degree of homology and/oroverlap (e.g., stringency). “Clustering” may be performed with thesequence data. For instance, a biomolecular sequence thought to beassociated with a particular molecular or biological activity in onetissue might be compared against another library or database ofsequences. This type of search is useful to look for homologous, andpresumably functionally related, sequences in other tissues or samples,and may be used to streamline the methods of the present invention inthat clustering may be used within one or more of the databases tocluster biomolecular sequences prior to performing a method of theinvention. The sequences showing sufficient homology with therepresentative sequence are considered part of a “cluster.” Such“sufficient” homology may vary within the needs of one skilled in theart.

[0678] As used herein, the term “internal database” refers to a databasemaintained within a local computer network. It contains, for example,biomolecular sequences associated with a project. It may also containinformation associated with sequences including, but not limited to, alibrary in which a given sequence is found and descriptive informationabout a likely gene associated with the sequence. The internal databasemay typically be maintained as a private database behind a firewallwithin an enterprise network. However, the invention is not limited toonly this embodiment and an internal database could be made available tothe public. The internal database may include sequence data generated bythe same enterprise that maintains the database, and may also includesequence data obtained from external sources.

[0679] The term “external database,” as understood herein, refers to adatabase located outside all internal databases. Typically, anenterprise network differing from the enterprise network maintaining theinternal database will maintain an external database. The externaldatabase may be used, for example, to provide some descriptiveinformation on biomolecular sequences stored in the internal database.In one embodiment, the external database is GenBank and associateddatabases maintained by the National Center for BiotechnologyInformation (NCBI), part of the National Library of Medicine.

[0680] As used herein and in the appended claims, the singular forms“a,” “an,” and “the” include plural reference unless the context clearlyindicates otherwise. Thus, for example, reference to a “compound” is areference to one or more such compounds and includes equivalents thereofknown to those skilled in the art, and so forth.

[0681] Unless defined otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood to one of ordinaryskill in the art to which this invention belongs. Although any methods,devices, and materials similar or equivalent to those described hereincan be used in the practice or testing of the invention, the preferredmethods, devices and materials are now described.

[0682] All publications and patents mentioned herein are incorporatedherein by reference for the purpose of describing and disclosing, forexample, the constructs and methodologies that are described in thepublications, which might be used in connection with the presentlydescribed invention. The publications discussed above and throughout thetext are provided solely for their disclosure prior to the filing dateof the present application. Nothing herein is to be construed as anadmission that the inventors are not entitled to antedate suchdisclosure by virtue of prior invention.

[0683] It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, constructs, and reagentsdescribed herein and as such may vary. It is also to be understood thatthe terminology used herein is for the purpose of describing particularembodiments only, and is not intended to limit the scope of the presentinvention which will be limited only by the appended claims.

EXAMPLES

[0684] The present invention is further illustrated by the followingexamples, which are not to be construed in any way as imposinglimitations upon the scope thereof, but rather are illustrative only. Onthe contrary, it is to be clearly understood that resort may be had tovarious other embodiments, modifications, and equivalents thereof which,after reading the description herein, may suggest themselves to one ofordinary skill in the art without departing from the spirit of thepresent invention or the scope of the appended claims.

[0685] The following acronyms, abbreviations, terms and definitions havebeen used throughout the experimental section. Acronyms orabbreviations: DIEA (N,N-diisopropylethylamine), THF (tetrahydrofuran),HPLC (high performance liquid chromatography), TLC (thin layerchromatography), mp (melting point), rt (room temperature), aq(aqueous), min (minute), h (hr, hour), atm (atmosphere), conc.(concentrated), MS (mass spectroscopy/spectrometry), NMR (nuclearmagnetic resonance), R_(f) (TLC retention factor), and R_(t) (HPLCretention time). NMR abbreviations: br (broad), apt (apparent), s(singlet), d (doublet), t (triplet), q (quartet), dq (doublet ofquartets), dd (doublet of doublets), dt (doublet of triplets), m(multiplet).

Example 1

[0686] General Synthetic, Purification, Characterization, andSpectroscopic Procedures

[0687] General Synthetic Procedures. Room temperature is defined as anambient temperature range, typically 20-25° C. An ice bath (crushedice/water) temperature is defined as a range, typically −5 to 0° C.Temperature at reflux is defined as ±15° C. of the boiling point of theprimary reaction solvent. Overnight is defined as a time range of 8-16hours. Vacuum filtration (water aspirator) is defined as range of 5-15mm Hg. Dried under vacuum is defined as using a high vacuum pump as arange of 0.1-5 mm Hg. Neutralization is defined as a typical acid-basedneutralization method and measured to a pH 6-8 range using pH-indicatingpaper. Brine is defined as a saturated aqueous sodium chloride. Nitrogenatmosphere is defined as positive static pressure of nitrogen gas passedthrough a Drierite column with an oil bubbler system. Concentratedammonium hydroxide is defined as an approximately 15 M solution.

[0688] All eluents for column or thin layer chromatography were preparedand reported as volume:volume (v:v) solutions, and HPLC eluent ratiosare v:v ratios. Aqueous sodium hydroxide or sodium bicarbonate solutionswere prepared as weight:volume (w:v) ratios. Aqueous hydrochloric acidsolutions were prepared as v:v ratios.

[0689] The quantities of solvents and/or reagents used for reactionwork-up or product isolation are those typically used by one trained inthe art of organic chemical synthesis, and the quantity of thesesolvents and/or reagents used is determined based upon syntheticexperience and appropriateness to the specific reaction. For example: 1)crushed ice quantity ranged from about 10-1000 g depending on reactionscale, 2) silica gel quantity used in column chromatography depended onmaterial quantity, complexity of mixture, and size of chromatographycolumn employed and ranged from about 5-1000 g, 3) extraction solventvolume ranged from about 10-500 mL depending on reaction size, 4) washesemployed in compound isolation ranged from about 10-100 mL of solvent oraq reagent depending on scale of reaction 5) drying reagents (potassiumcarbonate, sodium carbonate or magnesium sulfate) ranged from about5-100 g depending on the amount of solvent to be dried and its watercontent.

[0690] Melting points were measured against a mercury thermometer andare not corrected.

[0691] For column chromatography employing concentrated ammoniumhydroxide as part of the mobile phase, the fractions collected from thecolumn were dried over sodium sulfate, potassium carbonate or a mixtureof both. Then the organic layer was filtered by gravity or vacuum toremove the drying agent prior to concentration/evaporation.

[0692] Flash Chromatography. In the Tables, “ISCO” indicatespurification by flash chromatography as follows. Instrument: ISCOCombiFlashä Si 10×. Column: ISCO RediSepä—Disposable Columns for FlashChromatography (10 g of silica gel—normal phase—35-60 micron particlesize (230-400 mesh)). Mobile Phase A: CH₂Cl₂; Mobile Phase B: 10% NH₄OHin MeOH; Gradient: 0-10% B in 22 min, hold 10% B for 18 min; Fractions:30 fractions collected per column, 1.5 min each. Flow rate: 8.93 mL/min.The salient fractions were analyzed by MS and TLC (90:9:1CH₂Cl₂:MeOH:NH₄OH—R_(f) range 0.15-0.45) and combined in barcoded, taredvials. The resulting solutions were sampled for LC/MS analysis,concentrated in vacuo and their masses and yields were determined astabulated in the Tables.

[0693] If no additional purification was carried out after completion ofthe Parallel Synthesis, this is indicated as “None” in Table 2.

[0694] Analytical HPLC Procedures. Analytical HPLC procedures werecarried out according in one of two specific methods, depending uponavailability of instrumentation and sample requirements, as follows.

[0695] HPLC Method A. Column: Thomson Inst. Co. 4.6×50 mm C18 5 μm 60 A;Mobile Phase A: H₂O with 0.1% TFA; Mobile Phase B: CH₃CN with 0.1% TFA;Detection: UV 254 nm. Gradient 1: ELSD12MG; 10-90% B in 10 min, hold 90%B for 5 min; Flow: 1.0 mL/min. Gradient 2: ELSD5MG; 15-100% B in 5 min,hold 100% B for 3 min; Flow—2.0 mL/min.

[0696] HPLC Method B. Column: Thomson Inst. Co. 21×50 mm C 18 5 μm 60 A;Mobile Phase A: H₂O with 0.1% TFA; Mobile Phase B: CH₃CN with 0.1% TFA;Detection: UV 254 nm. Gradient 1: MIC8MG; 0-100% B in 8 min, hold 100% Bfor 2 min; Flow: 0.5 mL/min. Gradient 2: MIC15MG; 10-90% B in 15 min,hold 90% B for 3 min; Flow: 0.5 mL/min.

[0697] Preparative HPLC Procedures. Preparative HPLC was carried asfollows. Instrument: Gilson; Column: Thomson Inst. Co. 21.5×150 mm C18 5μm 60 A; Mobile Phase A: H₂O; Mobile Phase B: CH₃CN; Gradient: 15-100% Bin 10 min, hold 100% B for 5 min; Flow rate: 22 mL/min; Detection: UV254 nm. The fractions containing the desired compounds were collected inbarcoded, tared vials, sampled for LC/MS analysis, concentrated in vacuoand their masses and yields were determined as shown in the Tables.

[0698] Spectroscopic and Other Instrumental Procedures.

[0699] NMR. The ¹H and ¹³C NMR spectra described herein were obtainedusing Varian INOVA600 (600 MHz), Varian UNITY600 (600 MHz), or Varian400 (400 MHz) spectrometers. Spectrometer field strength and NMR solventused for a particular sample are indicated in the Examples, or on anyNMR spectra actually shown as Figures. Typically, ¹H NMR chemical shiftsare reported as δ values in parts per million (ppm) downfield fromtetramethylsilane (TMS) (δ=0 ppm) as an internal standard, and ¹³C NMRchemical shifts are reported in ppm downfield from TMS and referencedwith respect to the CDCl₃ signal center line (δ=77.0 ppm). Solid orliquid samples were dissolved in an appropriate NMR solvent (CDCl₃ orDMSO-d₆), placed in a NMR sample tube, and data were collected accordingto the spectrometer instructional manuals. Most samples were analyzed inVariable Temperature mode, typically at about 55° C., though some datafor some samples were collected with the probe at ambient temperature.NMR data were processed using NUTS: NMR Utility Transform Software (LiteVersion-20011128) by Acorn NMR.

[0700] LC-MS. The Liquid Chromatography-Mass Spectrometry (LC-MS)instrumentation used to examine the compounds of the present inventionwas typically a quadrupole/time-of-flight mass spectrometer, withelectrospray ionization (ESI). For example, the typical LC-MSinstrumentation used was a Micromass Q-Tof using electrospray ionization(ESI). This instrument is a quadrupole/time-of-flight mass spectrometercapable of mass resolution up to m/z of about 7500. Samples wereintroduced in a direct injection mode by first dissolving and dilutingthe sample in methanol or acetonitrile and injecting the sample solutioninto the ESI source via a 10 μL loop Rheodyne injection valve. Thecarrier solvent was typically a mixture of 70% CH₃CN or MeOH and 30% H₂O(v:v), containing about 0.1% formic acid. Accurate mass analyses wereperformed in a similar fashion except for using a multipoint masscalibration with the same instrument under high mass resolutionconditions. Samples were spiked with an appropriate internal massreference compound, as known by one of ordinary skill, and analyzed asdescribed above.

Example 2

[0701] General Methods for Parallel Synthesis

[0702] Examples 3-5 describe the synthetic procedures for thepreparation of the “library” of N²,N⁴,N⁶-tris(amino)-1,3,5-triazineswhich was prepared based on the strategy of changing only one pendantamino group per synthesis, and based on the parent structure 95 shownbelow, where each compound in the library contains two of the pendantgroups in 95.

[0703] The library was divided into three subgroups, and all threesubgroups are presented in Table 2. Library I (compounds 1-50) includescompounds having unchanged cycloheptylamino and[(1-ethyl-2-pyrrolidinyl)methyl]amino substituents, with various groupsbeing permuted at the remaining triazine amino position, prepared byMethod A as presented in Example 3. Library II (compounds 51-75)includes compounds having unchanged[(1-ethyl-2-pyrrolidinyl)methyl]amino and(3-fluoro-4-methoxyphenyl)amino substituents, with various groups beingpermuted at the remaining triazine amino position, prepared by Method Bas presented in Example 4. Library III (Compounds 76-100) includescompounds having unchanged (3-fluoro-4-methoxyphenyl)amino andcycloheptylamino substituents, with various groups being permuted at theremaining triazine amino position, prepared by Method C as described inExample 5. Thus, the combination of the specific amines employedproduced a library of compounds of novel composition. The sequence inwhich each monomer is added to form the compounds of the library is alsopresented in Table 2, because the Monomer 1 amine is added first,Monomer 2 amine added second, and Monomer 3 amine is added third.

Example 3

[0704] Parallel Synthetic Method A, for Library I Compounds

[0705] The following reaction scheme presents the general reagents andconditions for parallel synthetic method A used for the compounds ofTable 2 which designate Method A.

[0706] Reagents and conditions: (a) ArNHR, DIEA, CH₃CN/1,4-dioxane, −11C., 1 h (b) cycloheptylamine, DIEA, CH₃CN/1,4-dioxane, rt, overnight (c)2-(aminomethyl)-1-ethylpyrrolidine, DIEA, CH₃CN/1,4-dioxane, 80 C., 15

[0707] A stock solution of cyanuric chloride (0.542 M) in 1,4-dioxanewas prepared and 1 mL of this solution (containing 100 mg or 0.542 mmol)was dispensed into each of 50 barcoded 40 mL vials. These solutions werecooled to about −11° C. (freezing) using a J-KEM block connected to acirculating cooler. Meanwhile, individual solutions of each aryl amineArNHR (specified as Monomer 1 in Table 2, 0.542 mmol) anddisopropylethylamine (DIEA) (77 mg/104 μL, 0.596 mmol) in 1 mL of CH₃CNwere prepared. (For HCl salts, 204 μL DIEA (approx. 2.1 equiv) wasused.) Over a period of about 1 h, the amine/DIEA solutions were addedto the corresponding frozen cyanuric chloride solutions, one by one,with swirling. The resulting solutions were then shaken at about −11° C.for about 1 h and the reaction block was allowed to warm to roomtemperature over the next hour. The resulting2-amino-4,6-dichlorotriazine solutions were carried to the next stepwithout purification.

[0708] A stock solution of cycloheptylamine (1.08 M) and DIEA (1.19 M)in CH₃CN was prepared and 0.5 mL (containing 61 mg/69 μL, 0.542 mmolamine and 77 mg /104 μL, 0.596 mmol DIEA) was dispensed into each of the40 mL vials from the first step. The vials were shaken on the J-KEMblock overnight at room temperature and placed in a freezer (about −14°C.) without purification until the next reaction.

[0709] A stock solution of 2-(aminomethyl)-1-ethylpyrrolidine (1.08 M)and DIEA (1.19 M) in CH₃CN was prepared and 0.5 mL (containing 69 mg/79μL, 0.542 mmol amine and 77 mg/104 μL, 0.596 mmol DIEA) was dispensedinto each of the 40 mL from the second step. The vials were then shakenon the J-KEM block at about 80° C. for about 15 h. The solutions werecooled to room temperature and taken to dryness in vacuo. The residueswere then extracted with ethyl acetate and the extract was washed withbrine. The aqueous layers were extracted a second time with ethylacetate and the combined organic layers were dried over Na₂SO₄ andpassed through a plug of Celite™ into barcoded, tared vials. Afterconcentration in vacuo, masses were determined and yields werecalculated, and the compounds were sampled for LC/MS analysis.

Example 4

[0710] Parallel Synthetic Method B, for Library II Compounds

[0711] The following reaction scheme presents the general reagents andconditions for parallel synthetic method B, used for the compounds ofTable 2 which designate Method B.

[0712] Reagents and conditions: (a) 3-fluoro-p-anisidine, DIEA,CH₃CN/1,4-dioxane, −20 C., 1 h (b) R₂NHR, DIEA, CH₃CN/1,4-dioxane, rt,overnight (c) 2-(aminomethyl)-1-ethylpyrrolidine, DIEA,CH₃CN/1,4-dioxane, 80 C., 15

[0713] In an oven-dried round bottom flask, a solution of cyanuricchloride (5.0 g, 27.1 mmol) in 1,4-dioxane (40 mL) was cooled tofreezing in a CH₃CN/dry ice bath. To this frozen solution was added 40mL of CH₃CN, followed by DIEA (3.85 g/5.19 mL, 29.8 mmol). A solution of3-fluoro-p-anisidine (3.83 g, 27.1 mmol) in 10 mL of CH₃CN was thenadded slowly via syringe. The reaction mixture was stirred at about −20°C. for about 1 h and allowed to warm to room temperature over about 1 h.The resulting 2-amino4,6-dichlorotriazine solution was carried to thenext step without purification.

[0714] Fifty mL (13.5 mmol) of the prepared(4,6-dichloro-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl) aminesolution was divided equally (2 mL or 0.54 mmol each) among 25 barcoded,40 mL scintillation vials. Individual solutions of each R₂NHR (where R₂amine indicates Monomer 2 in Table 2, 0.542 mmol) and DIEA (77 mg/104μL, 0.596 mmol) in 0.5 mL of CH₃CN were prepared and added to thecorrespondingly labeled 40 mL vials. The resulting solutions were shakenon the J-KEM block overnight at room temperature and then placed in afreezer (about −14° C.) without purification until the next reaction.

[0715] A stock solution of 2-(aminomethyl)-1-ethylpyrrolidine (1.08 M)and DIEA (1.19 M) in CH₃CN was prepared and 0.5 mL (containing 69 mg/79μL, 0.542 mmol amine and 77 mg/104 μL, 0.596 mmol DIEA) was dispensedinto each of the 40 mL vials from the second step. The vials were shakenon the J-KEM block at about 80° C. for about 15 h. The solutions werecooled to room temperature and concentrated in vacuo. The residues werethen extracted with ethyl acetate and the extract washed with brine. Theaqueous layers were extracted a second time with ethyl acetate and thecombined organic layers were dried over Na₂SO₄ and passed through a plugof Celite™ into barcoded, tared vials. After concentration in vacuo,masses were calculated and the compounds were sampled for LC/MSanalysis.

Example 5

[0716] Parallel Synthetic Method C, for Library III Compounds

[0717] The following reaction scheme presents the general reagents andconditions for parallel synthetic method C, used for the compounds ofTable 2 which designate Method C.

[0718] Reagents and conditions: (a) 3-fluoro-p-anisidine, DIEA,CH₃CN/1,4-dioxane, −20 C., 1 h (b) cycloheptylamine, DIEA,CH₃CN/1,4-dioxane, rt, overnight (c) R₃NHR, DIEA, CH₃CN/1,4-dioxane, 80C., 15

[0719] In an oven-dried round bottom flask, a solution of cyanuricchloride (5.0 g, 27.1 mmol) in 1,4-dioxane (40 mL) was cooled tofreezing in a CH₃CN/dry ice bath. To this frozen solution was added 40mL of CH₃CN, followed by DIEA (3.85 g/5.19 mL, 29.8 mmol). A solution of3-fluoro-p-anisidine (3.83 g, 27.1 mmol) in 10 mL of CH₃CN was thenadded slowly via syringe. The reaction mixture was stirred at about −20°C. for about 1 h and allowed to warm to room temperature over 1 h. Theresulting 2-amino-4,6-dichlorotriazine solution was carried to the nextstep without purification.

[0720] Fifty mL (13.5 mmol) of the prepared(4,6-dichloro-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl) aminesolution was treated with a solution of cycloheptylamine (1.53 g/1.73mL, 13.5 mmol) and DIEA (1.93 g/2.60 mL, 14.9 mmol) in CH₃CN (8 mL). Theresulting solution was stirred overnight at room temperature and carriedto the next step without purification.

[0721] The resulting6-chloro-N-cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diaminesolution (13.5 mmol) was diluted up to 62.5 mL with CH₃CN and dividedequally (2.5 mL or 0.54 mmol each) between 25 barcoded 40 mLscintillation vials. Individual solutions of each R₃NHR (where R₃ amineindicates Monomer 3 in Table 2, 0.542 mmol) and DIEA (77 mg/104 μL,0.596 mmol) in 0.5 mL of CH₃CN were prepared and added to thecorrespondingly labeled 40 mL vial. The resulting solutions were shakenon the J-KEM block at about 80° C. for about 15 h. The solutions werecooled to room temperature and concentrated in vacuo. The residues werethen extracted with ethyl acetate and the extract washed with brine.Each organic layer was dried over Na₂SO₄ and passed through a plug ofCelite™ into a barcoded, tared vial. After concentration in vacuo,masses were calculated and the compounds were sampled for LC/MSanalysis.

Example 6 Synthesis of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cyclohexylmethyl-[1,3,5]triazine-2,4-diamine(102)

[0722]

[0723] To a sample of 101 (0.3004 g, 1.0 mmol, prepared as indicatedherein) dissolved in acetone (4 mL) was added a solution ofcyclohexanemethylamine (0.13 mL, 1.0 mmol) in acetone (1 mL) followed byaddition of a NaOH solution (0.0448 g, 1.0 mmol dissolved in 1 mL ofH₂O). The reaction mixture was allowed to stir at reflux for about 3hours. The reaction mixture was then poured over crushed ice andneutralized with 10% HCl (aq) and 5% NaOH (aq). The resulting solid wascollected by vacuum filtration, washed with water and dried overnightunder vacuum to afford compound 102 (0.29 g, 76% recovery).

Example 7 Synthesis ofN-(3-Chloro-4-methoxy-phenyl)-N′-cyclohexylmethyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(103)

[0724] To a sample of 102 (0.286 g, 1.0 mmol) dissolved in 1,4-dioxane(4 mL) was added a solution of N-methyl-4(methylamino)piperidine (0.15mL, 1.0 mmol) in acetone (1 mL) followed by addition of a NaOH solution(0.0462 g, 1.0 mmol dissolved in 1 mL of H₂O). The reaction mixture wasallowed to stir at about 80° C. for about 2 hours. The reaction mixturewas poured over crushed ice and neutralized with 10% HCl (aq). Theresulting solid was collected by vacuum filtration, washed with waterand dried under vacuum overnight. Column chromatography (silica gel,96:3:1 dichloromethane:methanol:conc. ammonium hydroxide) yielded alight purple solid 103 (41 mg, 9%), mp 84° C.; HPLC: YMC Pack Pro C18,40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 12.7 min,97% purity); ¹H NMR (600 MHz, CDCl₃, 55° C.) δ 7.98 (s, 1H), 7.18 (S,1H), 6.85 (d, J=9 Hz, 1H), 6.58 (s, 1H), 4.89 (s, 1H), 4.58-4.62 (m,1H), 3.87 (s, 3H), 3.25 (t, J=6.6 Hz, 2H), 3.05 (s, 3H), 2.94 (d, J=11.4Hz, 2H), 2.31 (s, 3H), 2.15 (S, 2H), 1.86 (dq, J=12, 4.2 Hz, 3H),1.57-1.78 (m, 8H), 1.15-1.30 (m, 4H), 1.00 (dq, J=11.4, 3 Hz, 2H); MS(ESI): m/z 476 (37.7), 474 (M+H, 100), 410 (1.4).

Example 8 Synthesis of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-(1-propyl-butyl)-[1,3,5]triazine-2,4-diamine(104)

[0725]

[0726] To a sample of 101 (0.3062 g, 1.0 mmol) dissolved in acetone (4mL) was added a solution of 4-heptylamine (0.15 mL, 1.0 mmol) in acetone(1 mL) followed by addition of a NaOH solution (0.0410 g, 1 mmoldissolved in 1 mL of H₂O). The reaction mixture was allowed to stir at30-50° C. for about about 3 hours. 10 The reaction mixture was thenpoured over crushed ice and neutralized with 10% HCl (aq) and 5% NaOH(aq). The resulting solid was collected by vacuum filtration, washedwith water and dried overnight under vacuum to afford 104 (0.363 g, 94%recovery).

Example 9 Synthesis ofN-(3-Chloro-4-methoxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-(1-propyl-butyl)-[1,3,5]triazine-2,4,6-triamine(105)

[0727] To a sample of 104 (0.363 g, 1.0 mmol) dissolved in 1,4-dioxane(6 mL) was added a solution of N-methyl-4(methylamino)piperidine (0.15mL, 1.0 mmol) in acetone (1 mL) followed by addition of a NaOH solution(0.0414 g, 1.0 mmol dissolved in 1 mL of H₂O). The reaction mixture wasallowed to stir at about 80° C. for about about 2 hours. The reactionmixture was poured over crushed ice and neutralized with 10% HCl (aq).The resulting solid was collected by vacuum filtration, washed withwater and dried under vacuum overnight. Column chromatography (silicagel, 96:3:1 dichloromethane:methanol:conc. ammonium hydroxide) yieldedlight purple solid 105 (97 mg, 20%), mp 249° C. HPLC: YMC Pack Pro C18,40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 14.4 min,98% purity; MS (ESI): m/z 476 (M+H, 100), 412 (2.9), 366 (2.8), 239(1.9).

Example 10 Synthesis ofN-(3-Chloro-4-methoxy-phenyl)-N′-isopropyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(106)

[0728]

[0729] To a sample of 101 (0.6157 g, 2.0 mmol) dissolved in anhydrous1,4-dioxane (15 mL) was added a solution of isopropylamine (0.17 mL, 2.0mmol) in anhydrous acetonitrile (1 mL) followed by addition of aN,N-diisopropylethylamine (DIEA) (0.38 mL, 2.2 mmol) in anhydrousacetonitrile (1 mL). The reaction mixture was allowed to stir at roomtemperature overnight under nitrogen. To this mixture was added DIEA(0.38 mL, 2.2 mmol) in anhydrous acetonitrile (1 mL) followed byaddition of N-methyl-4(methylamino)piperidine (0.29 mL, 2.0 mmol) inanhydrous acetonitrile (1 mL). The reaction mixture was allowed to stirat reflux overnight under nitrogen. The reaction mixture was extracted 3times with ethyl acetate. The combined organic layers were washed onetime with brine solution and dried over anhydrous potassium carbonate.The organic layer was concentrated on a rotary evaporator and allowed todry overnight under vacuum. Column chromatography (silica gel, 93:6:1CH₂Cl₂: CH₃OH: conc. NH₄OH) yielded light brown solid 106 (271 mg, 32%);TLC (silica gel, 93:6:1 CH₂Cl₂: CH₃OH: conc. NH₄OH), R_(f) 0.28; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 4.4 min, 84.8% purity; MS (ESI): m/z 422 (26), 420 (M+H,71.2), 378 (4.2), 231 (100), 211 (40.4), 118 (5.4).

Example 11 Synthesis ofN²-(3-chloro-4-methoxy-phenyl)-N⁴-isopropyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine(107)

[0730] Compound 107 was isolated (0.159 g) as a by-product via columnchromatography (silica gel, 93:6:1 CH₂Cl₂: CH₃OH: conc. NH₄0H); mp 129°C.; TLC (silica gel, 93:6:1 CH₂Cl₂: CH₃OH: conc. NH₄OH), R_(f) 0.14;HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 4.4 min, 93.5% purity; MS (ESI): m/z 408 (17.2),406 (M+H, 46.6), 375 (18.5), 245 (11.9), 224 (100), 204 (13.4).

Example 12 Synthesis of5-{4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-pentan-1-ol(108)

[0731]

[0732] To a sample of 101 (1.5046 g, 5.0 mmol) dissolved in anhydrous1,4-dioxane (30 mL) was added a solution of 5-amino-1-pentanol (0.5067g, 5.0 mmol) in anhydrous acetonitrile (12 mL) followed by addition ofN,N-diisopropylethylamine (DIEA) (0.95 mL, 5.5 mmol) in anhydrousacetonitrile (2 mL). The reaction mixture was allowed to stir at roomtemperature overnight under nitrogen. To the reaction mixture was addedDIEA (0.95 mL, 5.5 mmol) in anhydrous acetonitrile (1 mL) followed byaddition of N-methyl-4(methylamino)piperidine (0.73 mL, 5.0 mmol) inanhydrous acetonitrile (1 mL). The reaction mixture was allowed to stirat reflux overnight under nitrogen. The reaction mixture was extracted 3times with ethyl acetate. The combined organic layers were washed onetime with brine solution and dried over anhydrous potassium carbonate.The organic layer was concentrated on a rotary evaporator and allowed todry overnight under vacuum. Column chromatography (silica gel, 90:9:1CH₂Cl₂: CH₃OH: conc. NH₄OH) yielded light brown solid 108 (300 mg, 13%);TLC (silica gel, 90:9:1 CH₂Cl₂: CH₃OH: conc. NH₄OH), R_(f) 0.22; HPLC:YMC Pack Pro C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264nm, R_(t) 3.5 min, 74.8% purity; MS (ESI): m/z 466 (24.2), 464 (M+H,71.5), 378 (5.2), 253 (4.5), 244 (20.5), 233 (100), 216 (33.3), 196(14.6), 118 (5.1).

Example 13 Synthesis of5-[4-(3-chloro-4-methoxy-phenylamino)-6-(methyl-piperidin-4-yl-amino)-1,3,5-triazin-2-ylamino]-pentan-1-ol(109)

[0733] Compound 109 was isolated as a by-product (0.820 g) via columnchromatography (silica gel, 90:9:1 CH₂Cl₂: CH₃OH: conc. NH₄OH), mp 101°C.; TLC (silica gel, 90:9:1 CH₂Cl₂: CH₃OH: conc. NH₄OH), R_(f) 0.08;HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH:CH₃CN], 264 nm, R_(t) 3.6 min, 95.3% purity; MS (ESI): m/z 452 (13), 450(M+H, 35.6), 419 (3.9), 267 (5.1), 246 (100), 226 (21.3), 209 (23.6),118(1.1).

Example 14 Synthesis ofN-Butyl-6-chloro-N′-(3-chloro-4-methoxy-phenyl)-N-propyl-[1,3,5]triazine-2,4-diamine(110)

[0734]

[0735] To a sample of 101 (1.5334 g, 5.0 mmol) dissolved in acetone (20mL) was added a solution of N-propyl-butylamine (0.77 mL, 5.0 mmol) inacetone (1 mL) followed by the addition of NaOH (2.0 mL, 2.5 N, 5.0mmol). The reaction mixture was allowed to stir at 30-35° C. for about 3hours under nitrogen. The reaction mixture was extracted 3 times withdichloromethane; the combined organic layers were washed with brinesolution and dried over potassium carbonate. The sample was concentratedon a rotary evaporator and the resulting oil was dried overnight undervacuum. Column chromatography (silica gel, 96:3:1dichloromethane:methanol:conc. ammonium hydroxide) yielded light brownsolid 110 (1.4 g, 77% recovery).

Example 15 Synthesis ofN-Butyl-N′-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-N-propyl-[1,3,5]triazine-2,4,6-triamine(111)

[0736] To a sample of 110 (1.323 g, 3.4 mmol) dissolved in 1,4-dioxane(25 mL) was added a solution of N-methyl-4(methylamino)piperidine (0.4mL, 3.4 mmol) in 1,4-dioxane (1 mL) followed by the addition of NaOH(1.4 mL, 2.5 N, 3.4 mmol). The reaction mixture was allowed to stir atreflux for about 2 hours under nitrogen. The reaction mixture wasextracted 3 times with dichloromethane; the combined organic layers werewashed with brine and dried over potassium carbonate. The sample wasconcentrated on the rotary evaporator and the resulting solid was driedovernight under vacuum. Column chromatography (silica gel, 90:9:1dichloromethane:methanol:conc. ammonium hydroxide) yielded light brownsolid compound 111 (527 mg, 33%), mp 68° C.; TLC (silica gel, 90:9:1CH₂Cl₂: CH₃OH: conc. NH₄OH), R_(f) 0.46; HPLC: ODS-3V C18, 40:30:30[KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 41.6 min, 90.8%purity); MS (ESI): m/z 476 (M+H, 28.5), 261 (20.2), 260 (52.8), 259(100), 239 (18.6), 239 (50.6).

Example 16 Synthesis ofN²-Butyl-N⁴-(3-chloro-4-methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-N²-propyl-1,3,5-triazine-2,4,6-triamine(112)

[0737] Compound 112 was isolated as a by-product via columnchromatography, an oil (0.112 g); TLC (silica gel, 90:9:1 CH₂Cl₂: CH₃OH:conc. NH₄OH), R_(f) 0.23; HPLC: ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH3.2): CH₃OH: CH₃CN], 265 nm, R_(t) 41.4 min, 97.8% purity); MS (ESI):m/z 464 (11.6), 462 (M+H, 28.9), 431 (15.6), 273 (12.7), 253 (58.8), 252(100), 232 (25.8), 157 (14.5).

Example 17 Synthesis of 2,4-Dichloro-6-cyclohexylmethoxy-[1,3,5]triazine(113)

[0738]

[0739] To cyanuric chloride (3.76 g, 20.0 mmol) dissolved in toluene (20mL) was added potassium bicarbonate (2.80 g, 20.0 mmol) and 18-crown-6(0.1614 g, 0.6 mmol) followed by dropwise addition of cyclohexylmethanol(2.5 mL, 20 mmol) in 15 mL of toluene (15 mL). The reaction mixture wasallowed to stir at reflux for about 18 hours under nitrogen. Thereaction mixture was passed through a plug of Celite and concentratedusing a rotary evaporator and dried over night under vacuum to give 113as an oil (5.212 g, 99% recovery).

Example 18 Synthesis of(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-fluoro-4-methoxy-phenyl)-amine(114)

[0740] To a sample of 113 (1.011 g, 3.8 mmol) dissolved in acetone (20mL) was added a solution of 3-fluoro-p-anisidine (0.541 g, 3.8 mmol) inacetone (10 mL) followed by addition of NaOH (1.52 mL, 2.5 N, 3.8 mmol)and water (3 mL). The reaction mixture was allowed to stir at reflux forabout 3 hours under nitrogen. The reaction mixture was extracted 3 timeswith dichloromethane; the combined organic layers were washed with brinesolution and dried over potassium carbonate. The sample was concentratedon a rotary evaporator and the resulting oil was dried overnight undervacuum. Column chromatography (silica gel, 70:30 hexanes:ethyl acetate)yielded light yellow solid compound 114 (0.581 g, 42%), mp 98° C.; TLC(silica gel, 30:70 ethyl acetate:hexanes), R_(f) 0.36; MS (ESI): m/z 369(39.1), 368 (22.1), 367 (M+H, 100), 273 (3.2), 271 (10.7).

Example 19 Synthesis of6-Cyclohexylmethoxy-N,N′-bis-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4-diamine(115)

[0741] Compound 115 was obtained as a by-product (0.159 g) via columnchromatography (silica gel, 70:30 hexanes:ethyl acetate), mp 181° C.;TLC (silica gel, 30:70 ethyl acetate:hexanes), R_(f) 0.17; MS (ESI): m/z472 (M+H, 100), 261 (1.5).

Example 20 Synthesis of6-Cyclohexylmethoxy-N-(1-ethyl-pyrrolidin-2-ylmethyl)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(116)

[0742] To a sample of 114 (0.3004 g, 0.82 mmol) dissolved in 1,4-dioxane(15 mL) was added a solution of 2-(aminomethyl)-1-ethylpyrrolidine (0.12mL, 0.82 mmol) in acetone (1 mL) followed by the addition of NaOH (0.33mL, 2.5 N, 0.82 mmol) and water (1 mL). The reaction mixture was allowedto stir at reflux for about 2 hours under nitrogen. The reaction mixturewas extracted 3 times with dichloromethane; the combined organic layerswere washed with brine and dried over potassium carbonate. The samplewas concentrated on the rotary evaporator and the resulting solid wasdried overnight under vacuum. Column chromatography (silica gel, 93:6:1dichloromethane:methanol:conc. ammonium hydroxide) yielded a lightyellow solid, compound 116 (226 mg, 60%), mp 59° C.; HPLC: InertsilODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm,R_(t) 10.5 min, 100% purity; ¹H NMR (600 MHz, CDCl₃, 55° C.) δ 7.65(broad resonance, rotamers, 1H), 7.07 (br d, J=7.8 Hz, 1H), 6.90 (t, J=9Hz, 1H) 6.84 (broad resonance, rotamers, 1H), 4.12 (s, 2H), 3.88 (S,3H), 1.02 (s, 1H), 2.26 (apt sextet, J=6.6 Hz, 1H), 2.19 (q, J=9 Hz,1H), 1.16-1.92 (m, 10H), 1.57 (s, 2H), 1.17-1.32 (m, 3H), 1.05-1.11 (m,4H); MS (ESI): m/z459 (M+H, 100), 363 (40.7), 223 (16.1), 202 (4.4), 138(1.2).

Example 21 Synthesis of(4-Chloro-6-cyclohexylmethoxy-[1,3,5]triazin-2-yl)-(3-chloro-4-methoxy-phenyl)-amine(117)

[0743]

[0744] To a sample of compound 113 (1.012 g, 3.8 mmol) dissolved inacetone (20 mL) was added a solution of 3-chloro-p-anisidine (0.605 g,3.8 mmol) in acetone (10 mL) followed by addition of NaOH (1.52 mL, 2.5N, 3.8 mmol) and water (3 mL). The reaction mixture was allowed to stirat reflux for about 3 hours under nitrogen. The reaction mixture wasextracted 3 times with dichloromethane; the combined organic layers werewashed with brine and dried over potassium carbonate. The sample wasconcentrated on a rotary evaporator and the resulting oil was driedovernight under vacuum. Column chromatography (silica gel, 70:30hexanes:ethyl acetate) yielded a light peach colored solid, compound 117(0.547 g, 38%), mp 114° C.; TLC (silica gel, 30:70 ethylacetate:hexanes), R_(f) 0.44; MS (ESI): m/z 385 (74.3), 384, (22.9), 383(M+H, 100), 287 (8.3).

Example 22 Synthesis ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-1,3,5-triazine-2,4-diamine(118)

[0745] Compound 118 was obtained as a by-product (0.178 g) via columnchromatography (silica gel, 70:30 hexanes:ethyl acetate), mp 188° C.;TLC (silica gel, 30:70 ethyl acetate:hexanes), R_(f) 0.22; MS (ESI): m/z504 (M+H, 100), 379 (1), 338 (1.3).

Example 23 Synthesis ofN-(3-Chloro-4-methoxy-phenyl)-6-cyclohexylmethoxy-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine(119)

[0746] To a sample of 117 (0.3007 g, 0.78 mmol) dissolved in 1,4-dioxane(15 mL) was added a solution of 2-(aminomethyl)-1-ethylpyrrolidine (0.11mL, 0.78 mmol) in acetone (1 mL) followed by the addition of NaOH (0.31mL, 2.5 N, 0.78 mmol) and water (1 mL). The reaction mixture was allowedto stir at reflux for about 2 hours under nitrogen. The reaction mixturewas extracted 3 times with dichloromethane; the combined organic layerswere washed with brine solution and dried over potassium carbonate. Thesample was concentrated on the rotary evaporator and the resulting solidwas dried overnight under vacuum. Column chromatography (silica gel,93:6:1 dichloromethane:methanol:conc. ammonium hydroxide) yielded lightyellow solid compound 119 (159 mg, 43%), mp 140° C. HPLC: InertsilODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm,R_(t) 15.2 min, 99.7% purity; MS (ESI): m/z 475 (M+H, 64.1), 379 (49.5),231 (48.6), 210 (100), 190 (3.2).

Example 24 Synthesis of6-Chloro-N,N″-bis-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(120)

[0747]

[0748] To a sample of 101 (3.0556 g, 10.0 mmol) dissolved in acetone (25mL) was added a solution of 3-chloro-p-anisidine (1.6050 g, 10.0 mmol)in acetone (10 mL) followed by addition of NaOH (4.0 mL, 2.5 N, 10.0mmol). The reaction mixture was allowed to stir at rt for about 3 hoursunder nitrogen. The reaction mixture was poured over crushed ice. Theresulting solid was collected by vacuum filtration, washed with waterand dried overnight under vacuum to give compound 120 (4.06 g, 95%), mp213° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH: CH₃CN], 264 nm, R_(t) 70.0 min, 97.1% purity MS (ESI): m/z 427(20.90), 426 (M+H, 99.6), 210 (100), 209 (22.2), 196 (55.3), 169 (25.4).

Example 25 Synthesis ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(4-methyl-cyclohexyl)-[1,3,5]triazine-2,4,6-triamine(121)

[0749] To a sample of compound 120 (1.5004 g, 3.5 mmol) dissolved in1,4-dioxane (20 mL) was added a solution ofN-methyl-4(methylamino)-piperidine (0.5 mL, 3.5 mmol) in 1,4-dioxane (1mL) followed by the addition of NaOH (1.4 mL, 2.5 N, 3.5 mmol). Thereaction mixture was allowed to stir at reflux for about 2 hours undernitrogen. The reaction mixture was poured over crushed ice andneutralized with 10% HCl (aq). The resulting solid was collected byvacuum filtration, washed with water and dried overnight under vacuum.Column chromatography (silica gel, 96:3:1 dichloromethane:methanol:conc.ammonium hydroxide) yielded a purple solid, compound 121 (487 mg, 27%),mp 130° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 8.1 min, 96% purity; ¹H NMR (600 MHz,CDCl₃, 55° C.) δ 7.81-7.92 (broad resonance, 2H), 7.19-7.30 (broadresonance, 2H), 6.87 (d, J=9 Hz, 2H), 6.72 (s, 2H), 4.60-4.65 (m, 1H),3.88 (s, 6H), 3.05 (s, 3H), 2.95 (d, J=12 Hz, 2H), 2.32 (s, 3H), 2.19(t, J=11.4 Hz, 2H), 1.89 (dq, J=12.6, 3.6 Hz, 2H), 1.71 (apt d, J=11.4Hz, 2H), 1.65 (s, 1H); MS (ESI): m/z 519 (28.3), 518 (M+H, 42.1), 261(71.9), 260 (100).

Example 26 Synthesis ofN,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine(122)

[0750]

[0751] To a sample of 120 (1.5004 g, 3.5 mmol) dissolved in acetone (20mL) was added a solution of cycloheptylamine (0.4 mL, 3.5 mmol) inacetone (1 mL) followed by the addition of NaOH (1.4 mL, 2.5 N, 3.5mmol). The reaction mixture was allowed to stir at reflux for about 2hours under nitrogen. The reaction mixture was poured over crushed iceand neutralized with 10% HCl (aq). The resulting solid was collected byvacuum filtration, washed with water and dried overnight under vacuum togive light purple solid compound 122 (1.5 g, 85%), mp 183° C.; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 59 min, 96% purity; MS (ESI): m/z 503 (M+H, 29), 502(100), 458 (24.2), 425 (17.9), 225 (5.7), 155 (11.3), 114 (27.6).

Example 27 Synthesis ofN-(3-Bromo-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(123)

[0752]

[0753] To cyanuric chloride (0.184 g, 1.0 mmol) dissolved inacetonitrile (3 mL) stirring at about −10° C., was added a solution of3-bromo-p-anisidine (0.2019 g, 1.0 mmol) in acetonitrile followed by theaddition of N,N-diisopropylethylamine (DIEA) (0.17 mL, 1.0 mmol) inacetonitrile. The reaction mixture was allowed to stir at about −10° C.for 1 hour under nitrogen. The reaction mixture was then warmed to roomtemperature and allowed to stir at room temperature for another hourunder nitrogen. To the reaction mixture was added a solution ofcycloheptylamine (0.13 mL, 1.0 mmol) in acetonitrile followed byaddition of DIEA (0.17 mL, 1.0 mmol). The reaction mixture was allowedto stir at reflux overnight under nitrogen. To the reaction mixture wasadded N-methyl-4(methylamino)piperidine (0.13 mL, 1.0 mmol) inacetonitrile followed by the addition of DIEA (0.17 mL, 1.0 mmol). Thereaction mixture was allowed to stir at reflux for overnight undernitrogen. The reaction mixture was extracted 3 times with ethyl acetate;the combined organic layers were washed with brine solution and driedover potassium carbonate. The sample was concentrated on the rotaryevaporator and the resulting solid was dried overnight under vacuum.Column chromatography (silica gel, 90:9:1 methylene chloride:methanol:conc. ammonium hydroxide yielded 0.029 g (6%)of 123. ¹H NMR(400 MHz, CDCl₃) δ 7.97-8.19 (broad resonance, 1H), 7.12 (broadresonance, 1H), 6.78-6.80(m, 2H), 4.82 (br s, 1H), 4.58 (br s, 1H), 3.92(br s, 1H), 3.84 (s, 3H), 2.90-2.98 (m, 5H), 2.29 (s, 3H), 2.17 (broadresonance, 2H), 1.99-2.24 (broad resonance, 4H), 1.72-1.85 (m, 3H),1.42-1.62 (m, 11H); MS (ESI): m/z 520 (100), 518 (93.9), 458 (10.4), 424(20.8), 422 (21.1), 261 (67.5), 260 (63.4), 213 (13.9), 212 (13.6).

Example 28 Synthesis of6-Chloro-N-cyclohexylmethyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(125)

[0754]

[0755] To a sample of 124 (40.02 g, 138.4 mmol, prepared as indicatedherein) dissolved in acetone (300 mL) was added a solution ofcyclohexanemethylamine (18.0 mL, 138.4 mmol) in acetone (30 mL) followedby addition of NaOH (55.4 mL, 2.5 N, 138.4 mmol) and 130 mL of water.The reaction mixture was allowed to stir at reflux for about 3 hours.The reaction mixture was then poured over crushed ice and neutralizedwith 10% HCl (aq) and 10% NaOH (aq). The resulting solid was collectedby vacuum filtration, washed with water and dried overnight undervacuum. Recrystallization from ethyl acetate yielded a light yellowsolid, compound 125 (32.93 g, 65%), mp 156° C.; HPLC: Inertsil ODS-3VC18, 40:10:50 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 47.9min, 92% purity; MS (ESI): m/z 366 (M+H, 100).

Example 29 Synthesis ofN-Cyclohexylmethyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine(126)

[0756] To a sample of 125 (10.02 g, 27.3 mmol) dissolved in 1,4-dioxane(150 mL) was added a solution of 2-(aminomethyl)-1-ethylpyrrolidine (4.0mL, 27.3 mmol) in acetone (10 mL) followed by addition of NaOH (11 mL,2.5 N, 27.3 mmol) and 27 mL of water. The reaction mixture was allowedto stir at reflux for about 2 hours. The reaction mixture was extracted3 times with dichloromethane; the combined organic layers were washedwith brine and dried over potassium carbonate. The sample wasconcentrated on the rotary evaporator and the resulting solid was driedovernight under vacuum. Column chromatography (silica gel, 93:6:1dichloromethane:methanol:conc. ammonium hydroxide) yielded a lightyellow solid, compound 126 (7.014 g, 56%), mp 72° C.; HPLC: InertsilODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm,R_(t) 8.5 min, 93.4% purity; MS (ESI): m/z 458 (M+H, 37.3), 362 (4), 250(100), 230 (15.3), 229 (44.1).

Example 30 Synthesis ofN-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-pyrrolidin-1-yl-[1,3,5]triazine-2,4-diamine(128)

[0757]

[0758] To a sample of compound 127 (13.24 g, 36.2 mmol, prepared asindicated herein) dissolved in THF (150 mL) was added a solution ofpyrrolidine (3.0 mL, 36.2 mmol) in THF (10 mL) followed by addition ofNaOH (14.5 mL, 2.5 N, 36.2 mmol) and 36 mL of water. The reactionmixture was allowed to stir at reflux for about 2.5 hours. The reactionmixture was extracted 3 times with dichloromethane; the combined organiclayers were washed with brine and dried over potassium carbonate. Thesample was concentrated on the rotary evaporator and the resulting solidwas dried overnight under vacuum. Column chromatography (silica gel,98:2 dichloromethane:methanol) yielded light yellow solid 128 (3.36 g,23%), mp 79° C.; HPLC: Inertsil ODS-3V C18, 40:10:50 [KH₂PO₄ (0.01M, pH3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 24.5 min, 95.5% purity; ¹H NMR (600MHz, CDCl₃, 55° C.) δ 7.77 (broad resonance, 1H), 7.01-7.03 (m, 11H),6.86 (t, J=9 Hz, 1H), 6.62 (s, 1H), 4.80 (s, 1H), 4.02-4.06 (m, 1H),3.85 (s, 3H), 3.54 (s, 4 H), 1.99-2.03 (m, 2H), 1.91-1.93 (m, 3H),1.47-1.66 (m, 11H); MS (ESI): m/z 402 (30.7), 401 (M+H, 100).

Example 31 Synthesis of(4,6-Dichloro-[1,3,5]triazin-2-yl)-(3fluoro-4-methoxy-phenyl)-amine(124)

[0759]

[0760] To cyanuric chloride (28.84 g, 156.0 mmol) dissolved in acetone(200 mL) stirring at approximately 0-5° C., was added a solution of3-fluoro-p-anisidine (22.16 g, 156.0 mmol) in acetone (200 mL) followedby the addition of NaOH (63 mL, 2.5 N, 156.0 mmol). The reaction mixturewas allowed to stir at approximately 0-5° C. for about 2 hours. Thereaction mixture was then poured over crushed ice and neutralized with10% HCl (aq) and 5% NaOH (aq). The resulting solid was collected byvacuum filtration, washed with water and dried overnight under vacuum.Column chromatography (silica gel, 70:30 hexane:ethyl acetate) yieldedlight yellow solid compound 124 (29.6 g, 66%); mp 134° C.; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 20.3 min, 97.7% purity.

Example 32 Synthesis of6-Chloro-N-cycloheheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(127)

[0761] To a sample of 124 (10.00 g, 34.6 mmol) dissolved in acetone (150mL) was added a solution of cycloheptylamine (4.4 mL, 34.6 mmol) inacetone (20 mL) followed by addition of NaOH (13.8 mL, 2.5 N, 34.6 mmol)and 35 mL of water. The reaction mixture was allowed to stir at refluxfor about 3 hours. The reaction mixture was extracted 3 times withdichloromethane; the combined organic layers were washed with brine anddried over potassium carbonate. The sample was concentrated on therotary evaporator and the resulting solid was dried overnight undervacuum affording 127 (12.4 g, 98% recovery), mp 145° C.; HPLC: InertsilODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm,R_(t) 104.8 min, 97.3 % purity; ¹H NMR (600 MHz, CDCl3, 55° C.) δ7.50-7.64 (m, 1H), 7.02-7.03 (br resonance, 2H), 6.90 (t, J=8.9 Hz, 1H),5.35-5.41 (br resonance, 1H), 3.99 (br s, 1H), 4.12 (rotamer), 3.87 (s,3H), 2.01 (br s, 2H), 1.42-1.67 (m, 11H).

Example 33 Synthesis ofN-Cycloheptyl-N′-ethyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(129)

[0762] To 127 (11.00 g, 30 mmol) dissolved in THF (150 mL) was added asolution of ethylamine hydrochloride (2.43 mL, 30 mmol) in THF (20 mL)followed by addition of NaOH (24 mL, 2.5 N, 60 mmol) and 30 mL of water.The reaction mixture was allowed to stir at reflux for about 2 hours.The reaction mixture was extracted 3 times with dichloromethane; thecombined organic layers were washed with brine and dried over potassiumcarbonate. The sample was concentrated on the rotary evaporator and theresulting solid was dried overnight under vacuum. Column chromatography(silica gel, 98:2 dichloromethane:methanol) yielded a light yellow solid129 (4.81 g, 43%), mp 84° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 30.7 min, 94.2 %purity; ¹H NMR (600 MHz, CDCl₃, 55° C.) δ 7.69 (s, 1H), 7.00 (br d,J=7.0 Hz, 1H), 6.86 (t, J=8.4 Hz, 1H), 6.64 (s, 1H), 4.79-4.83 (brresonance, 2H), 4.01-4.03 (m, 1H), 3.85 (s, 3H), 3.38-3.42 (m, 2H),1.99-2.01 (m, 2H), 1.47-1.67 (m, 11H), 1.19 (t, J=7.2 Hz, 3H); MS (ESI):m/z 376 (29.5), 375 (M+H, 100).

Example 34 Synthesis ofN-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine(130)

[0763]

[0764] To 127 (5.009 g, 13.7 mmol) dissolved in THF (80 mL) was added asolution of 2-(aminomethyl)-1-ethylpyrrolidine (2.0 mL, 13.7 mmol) inTUF (10 mL) followed by addition of NaOH (5.5 mL, 2.5 N, 13.7 mmol) and13 mL of water. The reaction mixture was allowed to stir at reflux forabout 2 hours under N₂ atm. The reaction mixture was extracted 3 timeswith dichloromethane; the combined organic layers were washed with brineand dried over potassium carbonate. The sample was concentrated on therotary evaporator and the resulting solid was dried overnight undervacuum. Column chromatography (silica gel, 90:9:1 dichloromethane:methanol:conc. ammonium hydroxide) yielded a light yellow solid 130(3.63 g, 58%), mp 76° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄(0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 7.1 min, 97.1 % purity; MS(ESI): m/z 459 (16.5), 458 (M+H, 48.7), 362 (31.3), 250 (100), 230(22.8), 229 (62.7), 222 (17.2), 202 (34).

Example 35 Synthesis of2-[4-chloro-6-(3-chloro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-propane-1,3-diol(131)

[0765]

[0766] To 101 (0.6114 g, 2 mmol) dissolved in acetone (3 mL) was added2-amino-propane-1,3-diol (0.1818 g, 2 mmol) dissolved in acetone (1 mL)and water (1 mL). Then water (1 mL) was added to the reaction mixturefollowed by 2.5 N NaOH (aq) (0.8 mL, 2 mmol). The reaction mixture washeated at reflux for 3 h under a N₂ atmosphere. The reaction mixture wasdiluted with ethyl acetate and washed 2×brine. The organic layer wasseparated, dried over anhydrous K₂CO₃, filtered, and concentrated underreduced pressure affording 0.634 g of a purple solid. The crude materialwas purified by silica gel flash column chromatography eluting with 100%ethyl acetate affording a colorless oil 131 (0.124 g, 18%); HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 5.7 min, 83.3% purity; MS (ESI): m/z 360 (M+H, 100), 338(10.7), 183 (10.3)

Example 36 Synthesis of2-{4-(3-chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-propane-1,3-diol(132)

[0767] To 131 (0.979 g, 0.271 mmol) dissolved in 3 mL 1,4-dioxane wasadded methyl-4-(methylamino)piperidine (0.05 mL, 0.34 mmol) dissolved in2 mL 1,4-dioxane followed by the addition of 2.5 N NaOH (aq) (0.11 mL,0.275 mmol). The mixture was heated at reflux for 3 h 45 min, cooled toabout room temperature, and then concentrated under reduced pressure.The resulting material was diluted with dichloromethane and filtered.The filtrate was then concentrated affording 56.5 mg of material. Thecrude material was purified by silica gel pipet column eluting with 100%methanol affording an white solid 132 (21.1 mg, 18%), mp 84° C.; MS(ESI): m/z 454 (34.7), 452 (M+H, 100), 422 (11.3), 248 (25.3), 247(51.3), 157 (60.3), 129 (27.5).

Example 37 Synthesis ofN-(1-benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]-2,4,6-triamine(134)

[0768]

[0769] To 133 (0.1252 g, 0.382 mmol, prepared as indicated herein)dissolved in 3 mL acetonitrile was added N,N-diisopropyl ethyl amine(DIEA) (0.07 mL, 0.382 mL) followed by 4-amino-1-benzylamine (0.07 mL,0.382 mmol). The mixture was refluxed overnight under a N₂ atmosphere.The reaction mixture was diluted with methylene chloride and washed withbrine. The organic layer was separated, dried over K₂CO₃, filtered andconcentrated under reduced pressure to afford 0.159 g of material. Thecrude material was purified by silica gel flash column chromatographyeluting with 96:3:1 methylene chloride:methanol:conc. ammonium hydroxideand the collected fractions were dried over potassium carbonate,filtered and then concentrated under reduced pressure to afford 77 mg ofproduct. A second column under similar conditions was completed toafford an additional 30 mg of material for a combined product 134 (103mg, 50%); HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH: CH₃CN], 264 nm, R_(t) 13.7 min, 97.7% purity; MS (ESI): m/z 538(15.4), 536 (38.2), 448 (19.3), 446 (49.3), 290 (41.4), 289 (84.6), 269(100), 247 (4.4).

Example 38 Synthesis ofN²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine(135)

[0770]

[0771] To 134 (0.0485 g, 0.0867 mmol) in 2 mL methanol was added 10%Pd/C (0.052 g) followed by ammonium formate (0.0646 g, 1.02 mmol). Themixture was heated at reflux for about 1.5 h under a N₂ atmosphere. Thecooled reaction mixture was filtered by vacuum through Celite with amethylene chloride rinsing, and the filtrate concentrated under reducedpressure to afford 36 mg of material. The crude material was purified bysilica gel flash chromatography eluting with 90:9:1 methylenechloride:methanol:conc. ammonium hydroxide, and the collected fractionswere dried over potassium carbonate, filtered and then concentratedunder reduced pressure to afford a solid 135 (20 mg, 51.8%), mp 167° C.;HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH:CH₃CN], 264 nm, R_(t) 4.6 min, 52,1% (another major peak at R_(t) 7.3min, 46.9%); MS (ESI): m/z 448 (4.4), 446 (12.5), 412 (22.7), 386 (2.3),265 (32.9), 248 (42.6), 244 (56.2), 228 (37.1), 227 (100), 207 (6.9).

Example 39 Synthesis ofN²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine(136)

[0772]

[0773] To 133 (0.1257 g, 0.382 mmol, prepared as indicated herein)dissolved in 3 mL acetonitrile was added DIEA (0.07 mL, 0.382 mL)followed by 2-(aminomethyl)-1-ethyl pyrrolidine (0.06 mL, 0.382 mmol).The mixture was refluxed overnight under a N₂ atmosphere. The reactionmixture was diluted with methylene chloride and washed with brine. Theorganic layer was separated, dried over K₂CO₃, filtered and concentratedunder reduced pressure to afford 0.143 g of material. The crude materialwas purified by silica gel flash column chromatography eluting with96:3:1 methylene chloride:methanol:conc. ammonium hydroxide and thecollected fractions dried over approximately 1:1 potassiumcarbonate/sodium sulfate, filtered and then concentrated under reducedpressure to afford 77 mg of product. A second column under similarconditions was completed to afford an additional 30 mg of material for acombined 98 mg (54%) of a yellow colored solid 136, mp 69-70° C.; HPLC:YMC Pack Pro C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN], 264nm, R_(t) 12.9 min, 96.5% purity; MS (ESI): m/z 476 (16.3), 474 (42.9),260 (15), 259 (44.2), 258 (100), 238 (56), 216 (5.3), 210 (9.2).

Example 40 Synthesis of2-chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl-amino]-1,3,5-triazin-2-ylamino}-phenol(138)

[0774]

[0775] Under anhydrous conditions, 137 (0.1008 g, 0.21 mmol, prepared asdescribed herein) in a dry round bottomed flask was dissolved inanhydrous methylene chloride (3 mL) under a N₂ atmosphere about 0° C.(ice/water bath) was added BBr₃ (2.1 mL, 2.1 mmol, 1 M in methylenechloride) slowly by syringe. The mixture was stirred for about 2 hoursat about 0° C. and then quenched with water (5 mL). After standingovernight at rt, the mixture was diluted with ethyl acetate, water and10% NaHCO₃ (aq), and the organic layer was separated then washed withbrine. The organic layer was then dried over anhydrous sodium sulfate,filtered, and concentrated under reduced pressure to afford 0.648 g ofmaterial. The crude material was purified using silica gel flash columnchromatography eluting with 100% methanol to afford of a white solid 138(7 mg, 7%); HPLC: Inertsil ODS-3V C18, 40:30:30 [KE₂PO₄ (0.01 M, pH3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 4.9 min, 90.3% purity; ¹H NMR (600MHz, CDCl₃, 55° C.) (all resonances are broad) δ 7.93 (s, 1H), 7.13 (s,1H), 6.91-6.92 (m 1H), 6.55 (s, 1H), 4.80 (s, 1H), 4.59 (s, 1H), 4.02(s, 1H), 2.96-3.0 (m, 5H), 2.32 (s, 3H), 2.13 (s, 2H), 2.03 (s, 2H),1.86-1.88 (m, 2H), 1.53-1.67 (m, 12H); MS (ESI): m/z 463 (12.4), 461(27), 252 (59), 251 (100), 231 (32.3), 224 (1), 203 (9.8).

Example 41 Synthesis ofN²-cycloheptyl-N⁴—((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine(139)

[0776]

[0777] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atapproximately −10 to −20° C. was added 3-fluoro-p-anisidine (0.28 g, 2mmol) in CH₃CN followed by the addition of N,N-diisopropylethylamine(DIEA) (0.35 mL, 2 mmol) and stirred for an hour. The reaction mixturewas then allowed to reach room temperature for an hour. The second stepwas continued without further purification. Cycloheptylamine (0.25 mL, 2mmol) and DIEA (0.35 mL, 2 mmol) were added and the reaction mixture wasstirred overnight at rt. The third step was also preceded without anyfurther purification. S-(−)-2-aminomethyl-N-ethyl pyrrolidine (0.29 mL,2 mmol) and DIEA (0.35 mL, 2 mmol) were added and the reaction mixturewas refluxed overnight. The reaction mixture was diluted with ethylacetate and washed with brine. The organic layer was separated and driedover potassium carbonate, filtered, and concentrated under reducedpressure affording 0.920 g crude material. The crude material waspurified by column chromatography to yield a white solid 139 (0.550 g,60%), mp 75-77° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M,pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 7.9 min, 95.9% purity; MS (ESI):m/z 458 (M+H, 100).

Example 42 Synthesis ofN²-cycloheptyl-N⁴—((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine(140)

[0778]

[0779] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atabout −10 to −20° C. was added 3-fluoro-p-anisidine (0.28 g, 2 mmol) inCH₃CN followed by the addition of N,N-diisopropylethylamine (0.35 mL, 2mmol) and stirred for an hour. The reaction mixture was then allowed toreach room temperature for an hour. Then cycloheptylamine (0.25 mL, 2mmol) and DIEA (0.35 mL, 2 mmol) were added and the reaction mixture wasstirred overnight at rt. To this reaction mixtureR-(+)-2-aminomethyl-N-ethyl pyrrolidine (0.29 mL, 2 mmol) and DIEA (0.35mL, 2 mmol) were added and the reaction mixture was refluxed overnight.The reaction mixture was diluted with ethyl acetate and washed withbrine. The organic layer was separated and dried over potassiumcarbonate, filtered, and concentrated under reduced pressure affording0.920 g crude material. The crude material was purified by columnchromatography to yield a white solid 140 (0.500 g, 54.7%), mp 77-79°C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH:CH₃CN], 264 nm, R_(t) 7.9 min, 74.3% purity; MS (ESI): m/z 458 (M+H,100).

Example 43 Synthesis ofN²-cyclohexylmethyl-N⁴—((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxphenyl)-1,3,5-triazine-2,4,6-triamine(141)

[0780]

[0781] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atabout −20° C. was added 3-fluoro-p-anisidine (0.28 g, 2 mmol) in CH₃CNfollowed by the addition of N,N-diisopropylethylamine (DIEA) (0.35 mL, 2mmol) and stirred for about 1 hour. The reaction mixture was thenstirred at room temperature for about 1 hour. Then, cyclohexylmethylamine (0.26 mL, 2 mmol) and DIEA (0.35 mL, 2 mmol) were added and thereaction mixture was stirred overnight at RT. Then,S-(−)-2-aminomethyl-N-ethyl pyrrolidine (0.29 mL, 2 mmol) and DIEA (0.35mL, 2 mmol) were added and the reaction mixture was refluxed overnight.The reaction mixture was diluted with ethyl acetate and washed withbrine. The organic layer was separated and dried over sodium sulfate,filtered, and concentrated under reduced. The crude material waspurified by column chromatography eluting with 96:3:1 methylenechloride:methanol:conc. ammonium hydroxide to yield a white solid 141(0.400 g, 43.7%), mp 68-69° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 8.2 min, 97.1%purity; MS (ESI): m/z 458 (M+H, 100), 362 (2.8), 230 (85.4).

Example 44 Synthesis ofN²-cyclohexylmethyl-N⁴—((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine(142)

[0782]

[0783] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atabout −20° C. was added 3-fluoro-p-anisidine (0.28 g, 2 mmol) in CH₃CNfollowed by the addition of DIEA (0.35 mL, 2 mmol) and stirred for about1 hour. The reaction mixture was then stirred at room temperature forabout 1 hour. Then, cyclohexylmethyl amine (0.26 mL, 2 mmol) and DIEA(0.35 mL, 2 mmol) were added and the reaction mixture was stirredovernight at room temperature. Then, R-(+)-2-aminomethyl-N-ethylpyrrolidine (0.29 mL, 2 mmol) and DIEA (0.35 mL, 2 mmol) were added andthe reaction mixture was refluxed overnight. The reaction mixture wasdiluted with ethyl acetate and washed with brine. The organic layer wasseparated and dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The crude material was purified by columnchromatography eluting with 96:3:1 methylene chloride:methanol:conc.ammonium hydroxide to give 142 (0.100 g, 10.9%), mp 66-67° C.; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 8.2 min, 96.7% purity; ¹H NMR (600 MHz, CDCl₃) δ 7.58-7.73(broad resonance, 1H), 7.07-7.11 (broad resonance, 1H), 6.82 (t, J=9 Hz,1H), 5.49-5.65 (broad resonance, 1H), 4.96-5.13(broad resonance, 1H),3.82 (s, 3H), 3.54-3.70 (broad resonance, 1H), 3.13-3.20 (br m, 4H),2.81 (broad resonance, 1H), 2.54 (broad resonance, 1H), 2.05-2.18 (m,2H), 2.01 (s, 1H), 1.50-1.83 (brm, 9H), 1.05-1.22 (m, 5H), 0.91 (apt q,J=11.4 Hz, 2H); MS (ESI): m/z 458 (M+H, 100), 362 (3.8), 230 (99.8), 216(1), 182 (1.1).

Example 45 Synthesis of({4-cycloheptylamino-6-[((S)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile(143)

[0784]

[0785] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atabout −20° C. was added N-phenyl glycinonitrile (0.264 g, 2 mmol) inCH₃CN followed by the addition of DIEA (0.35 mL, 2 mmol) and stirred forabout 1 hour. The reaction mixture was then stirred at room temperaturefor about 1 hour. Then, cycloheptylamine (0.25 mL, 2 mmol) and DIEA(0.35 mL, 2 mmol) were added and the reaction mixture was stirredovernight at rt. Then, S-(−)-2-aminomethyl-N-ethyl pyrrolidine (0.29 mL,2 mmol) and DIEA (0.35 mL, 2 mmol) were added and the reaction mixturewas refluxed overnight. The reaction mixture was diluted with ethylacetate and washed with brine. The organic layer was separated and driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude material was purified by column chromatography eluting with96:3:1 methylene chloride:methanol:conc. ammonium hydroxide to yield143, (0.300 g, 33%) mp 53-55° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 6.9 min, 94.1%purity; MS (ESI): m/z 449 (M+H, 100), 381 (1.2), 353 (16.2), 226 (19.9),225 (54.3), 212 (20.5), 177 (18.3), 164 (9.6).

Example 46 Synthesis of({4-cycloheptylamino-6-[((R)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl)-phenyl-amino)-acetonitrie(144)

[0786]

[0787] To a mixture of cyanuric chloride (0.368 g, 2 mmol) in CH₃CN atabout −20° C. was added N-phenyl glycinonitrile (0.264 g, 2 mmol) inCH₃CN followed by the addition of DIEA (0.35 mL, 2 mmol) and stirred forabout 1 hour. The reaction mixture was then stirred at room temperaturefor about 1 hour. Then, cycloheptylamine (0.25 mL, 2 mmol) and DIEA(0.35 mL, 2 mmol) were added and the reaction mixture was stirredovernight at rt. Then, R-(+)-2-aminomethyl-N-ethyl pyrrolidine (0.29 mL,2 mmol) and DIEA (0.35 mL, 2 mmol) were added and the reaction mixturewas refluxed overnight. The reaction mixture was diluted with ethylacetate and washed with brine. The organic layer was separated and driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude material was purified by column chromatography eluting with96:3:1 methylene chloride:methanol:conc. ammonium hydroxide to yield144, (0.300 g, 33%), mp 53-55° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 6.8 min, 92.6%purity; MS (ESI): m/z 449 (M+H, 100), 381 (1.4), 353 (11.8), 226 (13),225 (33.1), 212 (15), 177 (13.5), 164 (7.8).

Example 47 Synthesis ofN²-[(1-ethyl-2-pyrrolidinyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-[(S)-2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5-triazine-2,4-diamine(145)

[0788]

[0789] Cyanuric chloride (11.07 g, 60 mmol) was dissolved in 40 mL CH₃CNand was cooled to about −20° C. To this was added DIEA (11.5 mL, 60mmol) followed by 3-fluoro-4-methoxyaninline (8.47 g, 60 mmol) in 20 mLCH₃CN (reaction froze). The reaction was allowed to warm to roomtemperature after about 1 hour at −20° C. TLC (2% CH₃OH/CH₂Cl₂) and massspectroscopy indicated the presence of the compound 124. The reactionmixture was cooled to about 0° C. before adding DIEA (11.5 mL, 66 mmol).2-Aminomethyl-1-ethylpyrrolidine (7.77 g, 60 mmol) in CH₃CN (10 mL) wasadded. The reaction was allowed to warm to rt and stirred overnight.Then DIEA (11.5 mL, 66 mmol) and S-(+)-2-methoxyethylpyrrolidine (6.91g, 60 mmol) in 20 mL 1,4-dioxane were added. The reaction was heated atabout 50° C. overnight. The solvent was removed in vacuo, and theresulting residue was purified by flash chromatography on silica gelpacked in ethyl acetate. The front running impurities were removed andsubsequently the eluent was increased in polarity to 10% CH₃OH: ethylacetate. The material collected from the column was then dissolved inwater and extracted in CH₂Cl₂ (4 times), dried over MgSO₄, andconcentrated to dryness to give a brown solid 145 (9.7 g, 27.6% yield),71-72° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2):CH₃OH: CH₃CN], 264 nm, R_(t) 5.37 min, 90.3 % purity; ¹H NMR (600 MHz,CDCl₃, 55° C.) δ 7.69 (s, 11H), 7.08 (d, J=7.8 Hz, 1H), 6.86 (t, J=9 Hz,1H), 4.29 (s, 1H), 3.90 -3.96 (m, 1H), 3.84 (s, 3H), 3.63-3.81 (m, 6H),3.35 (s, 3H), 3.23-3.25 (m, 1H), 2.85 (broad s, 1H), 2.78 (broad s 1H),2.14 (broad s, 2H), 1.89-2.04 (m, 6H), 1.37 (apparent t, J=7.2 Hz, 3H);¹³C NMR (150.8 MHz, CDCl₃, 55° C.) δ 165.8, 163.8 (2C), 152.3 (d,J_(c-f)=243.5 Hz), 143.0 (142.9, rotamer or diastereumer), 133.7(133.67, rotamer or diastereomer), 115.0, 114.4, 109.1 (108.9, rotameror diastereomer), 72.8, 66.6, 59.0, 57.0, 56.6, 53.7, 51.0, 46.8, 42.2,28.4 (28.2, rotamer or diastereomer), 23.1 (23.0, rotamer ordiastereomer), 10.9; MS (ESI) m/z 460.2 (M+H, 44.7), 251.1 (47.7), 235.1(27.5), 231.1 (37.4), 230.6 (100), 214.6 (36.5).

Example 48 Synthesis of(3-Chloro-4-methoxy-phenyl)-(4,6-dichloro-[1,3,5]triazin-2-yl)-amine(101)

[0790]

[0791] To cyanuric chloride (36.911 g, 200.0 mmol) dissolved in acetone(250 mL) stirring at approximately 0-5° C. (ice-water bath), was added asolution of 3-chloro-p-anisidine (31.528 g, 200.0 mmol) in acetone (150mL) followed by the addition of NaOH solution (80 mL, 2.5 N, 200.0mmol). The reaction mixture was allowed to stir at approximately 0-5° C.(ice-water bath) for about 1 hour. The reaction mixture was then pouredover crushed ice and neutralized with 10% HCl (aq). The resulting solidwas washed with water and dried overnight under vacuum to afford 101(58.3 g, 96%), mp 165° C.; HPLC: YMC Pack Pro C18, 40:30:30 [KH₂PO₄(0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 24.3 min, 97.8% purity);MS (ESI): m/z 305 (M+H, 100), 283 (26.3), 271 (26.9), 269 (75.2), 139(16.2).

Example 49 Synthesis of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(133)

[0792]

[0793] To a sample of compound 101 (20.02 g, 65.6 mmol) in acetone (200mL) was added cycloheptylamine (8.3 mL, 65.5 mmol) in acetone (55 mL)slowly by addition funnel at rt. Then water (66 mL) was added followedby aqueous sodium hydroxide (26.2 mL, 2. 5 N, 65.5 mmol) by additionfunnel. The reaction mixture was heated at reflux under a nitrogenatmosphere for approximately about 3 hours. The reaction was cooled,diluted with ethyl acetate, washed 1 time with water, and finally 1 timewith brine. The organic layer was separated and dried over potassiumcarbonate/sodium sulfate. The organic layer was filtered andconcentrated in vacuo. The product (24.13 g) was purified by flashcolumn chromatography (silica gel, 1:4 ethyl acetate: hexanes). Thefractions were combined and concentrated in vacuo to afford 133 as apale yellow solid (17.66 g, 70.5%), mp 146° C.; HPLC: Inertsil ODS-3VC18, 40:10:50 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 58.8min, 99.9% purity); MS (ESI): m/z 382 (M+H, 100), 241 (2.8), 226 (8.4),139 (43.5), 116 (6).

Example 50 Synthesis ofN-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(137)

[0794]

[0795] To 133 (10.014 g, 26.2 mmol) in 1,4-dioxane (80 mL) was addedslowly methyl-(1-methyl-piperidin-4-yl)-amine (3.8 mL, 26.2 mmol)dissolved in 1,4-dioxane (15 mL) by addition funnel. Then aqueous sodiumhydroxide (10.5 mL, 2.5 N, 26.2 mmol) was added by addition funnelfollowed by water (26 mL). The reaction mixture was heated at reflux forabout 2.5 hours under a nitrogen atmosphere. The reaction was cooled anddiluted with methylene chloride. The reaction mixture was filtered usingvacuum and the white solid 147 removed. The filtrate was then washed 1time with brine. The aqueous layer was back extracted 1 time withmethylene chloride. The organic layers were combined and dried overpotassium carbonate. The organic solution was filtered and concentratedin vacuo to afford the crude product (5.89 g). The crude reactionproduct was purified by flash column chromatography (silica gel) elutingwith 96:3:1 methylene chloride:methanol:15 M ammonium hydroxide. Thefractions were combined, dried over sodium sulfate/potassium carbonate,filtered, and concentrated in vacuo to afford 137 as a white solid (3.84g, 30.9%), mp 104-105° C.; HPLC: YMC Pack Pro C18, 40:30:30 [KH₂PO₄(0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 13.8 min, 97% purity); MS(ESI): m/z 474 (M+H, 41), 408 (2.3), 364 (2.8), 258 (13), 239 (14), 239(47.5), 238 (100), 127 (5.3).

Example 51 Synthesis ofN²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine(146)

[0796] Compound 146 was isolated as a by-product via columnchromatography (silica gel, 96:3:1 methylene chloride:methanol:conc.ammonium hydroxide, mp 114-116° C.; TLC (silica gel, 90: 9: 1, CH₂Cl₂:CH₃OH; conc. NH₄OH), R_(f) 137 0.31 and R_(f) 146 0.15; HPLC: InertsilODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm,R_(t) 10.7 min, 91.1% purity); MS (ESI): m/z 460 (M+H, 25.4), 364(17.9), 292 (2), 273 (17.1), 272 (37.9), 252 (44), 251 (100), 231 (2.2),157 (10.54), 118 (2.8).

Example 52 Synthesis of4-(3-Chloro-4-methoxy-phenylamino)-6-cycloheptylamino-1,3,5-triazin-2-ol(147)

[0797] Compound 147 was isolated as a by-product by vacuum filtrationprior to isolation of 137, white solid, mp >310° C.; MS (ESI); m/z 727([2(363)+H], 1.2, 364 (M+H, 100).

Example 53 Synthesis ofN-(1-Aza-bicyclo[2.2.2]oct-3-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-)1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4,6-triamine(148)

[0798]

[0799] To 101 (3.056 g, 10.0 mmol) dissolved in anhydrous acetonitrile(30 mL) at about 0° C. was added a solution of2-(aminomethyl)-1-ethylpyrrolidine (1.5 mL, 10.0 mmol) in anhydrousacetonitrile (5 mL) followed by addition of a DIEA (1.9 mL, 11.0 mmol).The reaction mixture was allowed to warm to room temperature and wasstirred at room temperature overnight under nitrogen. Then DIEA (1.9 mL,11 mmol) was added which was followed by addition of 3-aminoquinuclidinedihydrochloride (1.962 g, 10.0 mmol) in 1,4-dioxane (5 mL). The reactionmixture was allowed to stir at reflux overnight under nitrogen. Thereaction mixture was extracted 2 times with dichloromethane and 1 timewith ethyl acetate. The combined organic layers were washed one timewith brine and dried over anhydrous potassium carbonate. The organiclayer was with 20% HCl (aq). The aqueous layer was neutralized with 2.5N NaOH (aq) and then extracted 3 times with ethyl acetate. The combinedorganic layers were washed 1 time with brine, dried over potassiumcarbonate, concentrated on a rotary evaporator and allowed to dryovernight under vacuum. Column chromatography (silica gel, 85:14:1dichloromethane:methanol:conc. ammonium hydroxide) yielded a pale whitesolid 148 (100 mg, 2%), mp 83° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 8.1 min, 71.2%purity); MS (ESI): m/z 488 (M+H, 18.7), 280 (100), 245 ([M+2H]++, 37.4),236 (23.5).

Example 54 Synthesis ofN²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine(149)

[0800]

[0801] To a mixture of cyanuric chloride (1.8 g, 9.7 mmol) in CH₃CN atabout −20° C. was added 2-chloro-N,N-diethyl phenylene-1,4-diaminehydrochloride (2.35 g, 10 mmol) in CH₃CN followed by the addition ofN,N-diisopropylethylamine (DIEA) (1.75 mL, 10 mmol) and stirred for anhour. The reaction mixture was then allowed to reach room temperaturefor about 1 hour. Then cycloheptylamine (1.25 mL, 9.8 mmol) and DIEA(1.75 mL, 10 mmol) were added and the reaction mixture was stirredovernight at rt. Then, 2-(aminomethyl)-1-ethylpyrrolidine (1.45 mL, 10mmol) and DIEA (1.75 mL, 10 mmol) were added and the reaction mixturewas refluxed overnight. The reaction mixture was diluted with ethylacetate and washed with brine. The organic layer was separated and driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude material was purified by column chromatography (silica gel)eluting with 96:3:1 methylene chloride:methanol:conc. ammonium hydroxideto yield 149 (0.800 g, 15%) as a white solid, mp 84-85° C.; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2): CH₃OH: CH₃CN],264 nm, R_(t) 9.5 min, 96% purity; MS (ESI): m/z 515 (M+H, 9.4), 259(16.8), 258 (55.1), 257 (100).

Example 55 Synthesis ofN²-cycloheptyl-N⁴-(2-dimethylamino-ethyl)-N⁶-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine(150)

[0802]

[0803] Cyanuric chloride (1.84 g, 10 mmol) in CH₃CN (20 mL) was cooledto about −10° C. was added 3-fluoro-p-anisidine (1.41 g, 10 mmol)followed by DIEA (1.8 mL, 10 mmol). The reaction was stirred for about45 min then at room temperature for about 45 min under an N₂ atmosphere.Cycloheptylamine (1.26 mL, 10 mmol) was added followed by DIEA (1.8 mL,10 mmol) and the reaction was stirred at room temperature overnight.N,N-dimethylethylenediamine (1.1 mL, 10 mmol) was added followed by DIEA(1.8 mL, 10 mmol) and the mixture was heated at reflux under N₂overnight. The reaction was diluted with ethyl acetate, washed withbrine, and dried over anhydrous K₂CO₃. The material (1.178 g) waspurified by silica gel column chromatography to afford a solid 150(1.178 g, 28%), mp 73-76° C.; HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01M, pH 3.2): CH₃OH: CH₃CN], 264 nm, R_(t) 10.8 min, 95.1%purity; MS (ESI): m/z 418 (M+H, 100), 373 (11.9), 322 (7.8), 277 (6.8),162 (3.6).

Example 56 Synthesis of({4-cycloheptylamino-6-[1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile(151)

[0804]

[0805] To cyanuric chloride (1.84 g, 10 mmol) in CH₃CN (20 mL) at about−10 to −20° C. was added DIEA (1.75 mL, 10 mmol) and N-phenylglycinonitrile (1.3 g, 10 mmol), and stirred for about 1 hour. Thereaction mixture was then allowed to reach room temperature for an hour.To this reaction mixture, DIEA (1.75 mL, 10 mmol) and cycloheptylamine(1.25 mL, 10 mmol) were added and the reaction mixture was stirredovernight at rt. Then, DIEA (1.75 mL, 10 mmol) and2-aminomethyl-N-ethylpyrrolidine (1.45 mL, 10 mmol) were added and thereaction mixture was refluxed overnight. The reaction mixture wasworked-up, isolated, and then purified by column chromatography (silicagel) eluting with 96:3:1 methylene chloride:methanol:conc. ammoniumhydroxide to yield 151, (3 g, 66%), mp 52-54° C.; MS (ESI): m/z 449(M+H, 100), 225 [(M+2H)²⁺, 22.3].

Example 57 Synthesis ofN-Azepan-1-yl-6-chloro-N′-(3-chloro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(152)

[0806]

[0807] To 101 (6.03 g, 20.0 mmol) dissolved in acetone (75 mL) was addeda solution of 1-aminohomopiperidine (2.3 mL, 20.0 mmol) in acetone (10mL) followed by addition of NaOH (8.0 mL 2.5 N NaOH solution, 20.0 mmol)and 20 mL of water. The reaction mixture was allowed to stir at refluxovernight under nitrogen. The reaction mixture was extracted 3 timeswith dichloromethane; the combined organic layers were washed with brineand dried over potassium carbonate. The sample was concentrated on arotary evaporator and the resulting oil was dried overnight undervacuum. Column chromatography (96:3:1 dichloromethane:methanol:conc.ammonium hydroxide) yielded a light purple solid 152 (1.2 g, 16%), mp139° C.; TLC (silica gel, 96:3:1, CH₂Cl₂, CH₃OH, conc. NH₄OH), R_(f)0.31; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2):CH₃OH: CH₃CN], 264 nm, R_(t) 52.5 min, 94.9% purity; MS (ESI): m/z 383(M+H, 100).

Example 58 Synthesis ofN″-(3-chloro-4-methoxy-phenyl)-N,N′-bis-perhydro-azepin-1-yl-1,3,5-triazine-2,4,6-triamine(153)

[0808] Compound 153 was isolated as a by-product (2.3 g) by columnchromatography (silica gel, 96:3:1, CH₂Cl₂, CH₃OH, conc. NH₄OH), mp 199°C.; TLC (silica gel, 96:3:1, CH₂Cl₂, CH₃OH, conc. NH₄OH), R_(f) 0.11;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH 3.2): CH₃OH:CH₃CN], 264 nm, R_(t) 15 min, 86% purity); MS (ESI): m/z 461 (M+H, 100),366 (19.7), 365 (19.6), 232 (11), 231 (27.3).

Example 59 Synthesis ofN-Azepan-1-yl-N′-(3-chloro-4-methoxy-phenyl)-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(154)

[0809]

[0810] To 152 (0.2007 g, 0.5 mmol) dissolved in THF (10 mL) was added asolution of N-methyl-4(methylamino)piperidine (0.07 mL, 0.5 mmol) in THF(1 mL) followed by the addition of DIEA (1.0 mL, 0.55 mmol) inacetonitrile (1 mL). The reaction mixture was allowed to stir at refluxovernight under nitrogen. The reaction mixture was extracted 3 timeswith dichloromethane; the combined organic layers were washed with brineand dried over potassium carbonate. The sample was concentrated on arotary evaporator and the resulting oil was dried overnight undervacuum. Column chromatography (90:9:1 dichloromethane:methanol:conc.ammonium hydroxide) yielded a light yellow solid 154 (65 mg, 27%), mp100° C.; TLC (silica gel, 90:9:1 CH₂Cl₂: CH₃OH, conc. NH₄OH), R_(f)0.36; MS (ESI): m/z 475 (M+H, 23.2), 378 (11.6), 258 (68.9), 239 (52.2),238 (100).

Example 60 Synthesis ofN⁴-(3-chloro-4-methoxy-phenyl)-N⁶-methyl-N²-perhydro-azepin-1-yl-N⁶-piperidin-4-yl-1,3,5-triazine-2,46-triamine (155)

[0811] Compound 155 was obtained as a by product (50 mg) of the reactionvia column chromatography (silica gel, 90:9:1dichloromethane:methanol:conc. ammonium hydroxide), mp 81° C.; TLC(silica gel, 90:9:1 CH₂Cl₂:CH₃OH, conc. NH₄OH), R_(f) 0.25; MS (ESI):m/z 461 (M+H, 20.3), 430 (2.8), 273 (11.8), 272 (25.5), 251 (100), 236(4.6), 215 (4.7).

Example 61 Synthesis ofN,N′-di-n-propyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine(156)

[0812]

[0813] To cyanuric chloride (0.368 g, 2 mmol) in CH₃CN at about −20° C.was added 3-fluoro-p-anisidine (0.28 g, 2 mmol) in CH₃CN followed by theaddition of DIEA (0.39 mL, 2.2 mmol) and stirred for about 1 hour. Thereaction mixture was then stirred at room temperature for about 1 hour.Then n-propylamine (1.64 mL, 19.9 mmol) and DIEA (0.39 mL, 2.2 mmol)were added and the reaction mixture was stirred overnight at rt. Thereaction mixture was worked up as usual, diluted with ethyl acetate andwashed with brine. The organic layer was separated and dried over sodiumsulfate, filtered, concentrated under reduced pressure, and compound 156was purified by silica gel column chromatography. mp 53-55° C.; HPLC:Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264nm, R_(t) 12.6 min, 93.7% purity; MS (ESI): m/z 335 (M+H, 100), 331(1.5),126 (1).

Example 62 Synthesis ofN,N′-dicyclopropyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine(157)

[0814]

[0815] To cyanuric chloride (0.368 g, 2 mmol) in CH₃CN at about −20° C.was added 3-fluoro-p-anisidine (0.28 g, 2 mmol) in CH₃CN followed by theaddition of DIEA (0.39 mL, 2.2 mmol) and stirred for about I hour. Thereaction mixture was then stirred at room temperature for about 1 hour.Then cyclopropylamine (1.39 mL, 20 mmol) and DIEA (0.39 mL, 2.2 mmol)were added and the reaction mixture was stirred overnight at rt. Thereaction mixture was worked up as usual, diluted with ethyl acetate andwashed with brine. The organic layer was separated and dried over sodiumsulfate, filtered, concentrated under reduced pressure, and compound 157was purified by silica gel column chromatography (200 mg, 30%), mp91-92° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 8.6 min, 99.1% purity; MS (ESI): m/z331(M+H, 100), 305 (0.8), 151 (0.3).

Example 63 Synthesis ofN′-Cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine(158)

[0816]

[0817] To cyanuric chloride (0.180 g, 1 mmol) in 1,4-dioxane (1 mL) atabout −10 to −20° C. was added N,N-diisopropylethylamine (DIEA) (0.19mL, 1mmol) in CH₃CN (1 mL) and 3-fluoro-p-anisidine (0.14 g, 1 mmol) inCH₃CN (1 mL) and stirred for about 1 hour. The reaction mixture was thenstirred at room temperature for about 1 hour. Then a solution ofcycloheptylamine (0.13 mL, 1 mmol) and DIEA (0.19 mL, 1 mmol) in CH₃CN(0.5 mL) was added and the reaction mixture was stirred overnight at rt.Then, N-methyl-4(methylamino)piperidine (0.15 mL, 1 mmol) and DIEA (0.19mL, 1 mmol) in CH₃CN (0.5 mL) were added and the reaction mixture wasrefluxed overnight. The reaction mixture was worked-up using saturatedsodium bicarbonate, and brine. The organic layer was separated and driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude material was purified by column chromatography (silica gel,90:9:1 dichloromethane:methanol:conc. ammonium hydroxide) to give 158(0.130 g, 28%); TLC (silica gel, 90:9:1, CH₂Cl₂, CH₃OH, conc. NH₄OH),R_(f) 0.26); ¹H NMR (600 MHz, CDCl₃, 55° C.) δ 7.74 (br s, 1H), 6.94 (brs, 1H), 6.81-6.84 (m, 2H), 4.83 (br resonance, 1H), 4.55 (s, 1H), 3.98(s, 1H), 3.82 (s, 3H), 2.97 (s, 3H), 2.94 (br d, J=11.9 Hz, 2H), 2.29(s, 3H), 2.06-2.10 (m, 2H), 1.93-1.97 (m, 2H), 1.84-1.90 (m, 2H),1.44-1.66 (m, 12H).

Example 64 Synthesis ofN²-Cycloheptyl-N⁴-(3-fuoro-4methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine(159)

[0818] Compound 159 was isolated as a by-product (55 mg) by columnchromatography (silica gel, 90:9:1 dichloromethane:methanol:conc.ammonium hydroxide); TLC (silica gel, 90:9:1, CH₂Cl₂, CH₃OH, conc.NH₄OH), R_(f) 0.1);

[0819] HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 8.3 min, 93.5% purity; MS (ESI): m/z443 (M+H, 100).

Example 65 Synthesis ofN²-cycloheptyl-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine,hydrogen chloride salt (160)

[0820]

[0821] To 171 in dry methanol (1 mL, prepared according to parallelsynthesis Method C using the appropriate monomers, as disclosed herein)was added HCl (0.3 mL, 0.3 mmol, 1 M in diethyl ether) by syringe undera N₂ atmosphere. The mixture was stirred for 10 min at room temperature,concentrated and dried in vacuo overnight to give an off-white solid 160(0.131 g) that is water soluble, mp 189-190° C. (at 160° C. sample turnsbrown); HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 7.3 min, 89.1% purity.

Example 66 Synthesis of[N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]trizaine-2,4,6-triamine(161)

[0822]

[0823] To 137 (0.473 g, 1.0 mmol) dissolved in methanol (5 mL) was added1.0 M hydrochloric acid in diethyl ether (1.0 mL, 1 mmol). The reactionmixture was allowed to stir for about 1 hour at room temperature. Thereaction mixture was then concentrated on a rotary evaporator. Theresulting solid was dissolved in water, filtered and concentrated on therotary evaporator. The sample was freeze dried under vacuum and a solid161 (359.1 mg, 70%) was collected, mp 173-176° C.

Example 67 Synthesis ofN²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triaminehydrogen chloride salt (163)

[0824]

[0825] To 162 (1.0 g, 2 mmol, prepared according to parallel synthesismethod A with the appropriate monomers, as disclosed herein) in methanol(10 mL) was added HCl (2.5 mL, 2.5 mmol, 1 M) in diethyl ether andstirred. The reaction mixture was evaporated. It was then dissolved inwater, filtered, evaporated in vacuo, and dried over night under vacuumto afford a solid 163 (1.1 g, 93%).

Example 68 Synthesis ofN²-cycloheptyl-N⁴-(1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triaminehydrogen chloride salt (164)

[0826]

[0827] To 130 (2.285 g, 5 mmol) in dry methanol (10 mL) was added HCl (5mL, 5 mmol, 1 M in diethyl ether) and stirred at room temperature forabout 1 hour. The reaction was evaporated in vacuo, dissolved in water,filtered, evaporated and then dried under vacuum overnight to afford asolid 164 (2.396 g, 97%), mp 131-133° C.; HPLC: Inertsil ODS-3V C18,40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 7.9 min,98.2% purity.

Example 69 Synthesis ofN²-(cyclohexylmethyl)-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(4-fluoro-3-methoxyphenyl)-1,3,5-triazine-2,4,6-triaminehydrogen chloride salt (165)

[0828]

[0829] To 136 (0.457 g, 1 mmol) in dry diethyl ether was added HCl (1mL, 1 mmol, 1 M in diethyl ether). A precipitate formed immediately. Themixture was stirred at room temperature for about 1 hour, and thenconcentrated in vacuo. The resulting material was dissolved in water,filtered, evaporated, and dried overnight in vacuo to give a solid 165(0.400 g, 81%), mp 85° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄(0.01 M, pH 3.2):CH₃H:CH₃CN], 264 nm, R_(t) 8.2 min, 89.6% purity;

Example 70 Synthesis of({4-cycloheptylamino-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrilehydrogen chloride salt (166)

[0830]

[0831] To 151 (0.448 g, 1 mmol) in dry diethyl ether (2 mL) was addedHCl (1 mL, 1 mmol, 1 M in diethyl ether). The mixture was stirred atroom temperature for about 1 hour, and then concentrated in vacuo. Theresulting material was dissolved in water (5-10 mL), filtered,evaporated, and dried overnight under vacuum to give a solid 166 (0.418g, 86%), mp 125-127° C.; HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄(0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 6.9 min, 73.4% purity.

Example 71 Synthesis ofN²-cycloheptyl-N⁴-(3fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triaminemaleate salt (167)

[0832]

[0833] Compounds 158 (100.3 mg, 0.219 mmol) and maleic acid (25.4 mg,0.219 mmol) were dissolved in CH₃OH (2 mL) and stirred at roomtemperature under a N₂ atmosphere for about 75 min. The reaction mixturewas filtered through a cotton plug and concentrated in vacuo to afford asolid 167, 0.1239 g, mp 99-100° C. In a qualitative test, this materialwas water-soluble. HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M,pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 7.7 min, 87.9% purity.

Example 72 Synthesis ofN²-cycloheptyl-N⁴-(3fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triaminecitrate salt (168)

[0834]

[0835] Compounds 158 (100 mg, 0.219 mmol) and citric acid (42.1 mg,0.219 mmol) were dissolved in CH₃OH (2 mL) and stirred at roomtemperature under a N₂ atmosphere for about 2 hours. The reactionmixture was filtered through a cotton plug and concentrated in vacuo toafford a solid 168 (0.1387 g), mp 125° C. In a qualitative test, thismaterial was water insoluble. HPLC: Inertsil ODS-3V C18, 40:30:30[KH₂PO₄ (0.01M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 7.7 min, 90.1%purity.

Example 73 Synthesis ofN²-cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-(1-methylpiperidin-4-yl)-1,3,5-triazine-2,4,6-triamine succinate salt (169)

[0836]

[0837] Compounds 158 (101.5 mg, 0.219 mmol) and succinic acid (24.8 mg,0.219 mmol) were dissolved in CH₃OH (2 mL) and stirred at roomtemperature under a N₂ atmosphere for about 75 min. The reaction mixturewas filtered through a cotton plug and concentrated in vacuo to afford asolid 169 (0.1248 g), mp 81° C. In a qualitative test, this material waswater-soluble. HPLC: Inertsil ODS-3V C18, 40:30:30 [KH₂PO₄ (0.01M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 7.6 min, 89.8% purity.

Example 74 Synthesis ofN-(3-Bromo-4-methoxy-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triaminehydrogen chloride salt (170)

[0838] To 123 (1.0 mmol) dissolved in methanol (5 mL) was added 1.0 Mhydrochloric acid in diethyl ether (1.0 mL, 1 mmol). The reactionmixture was allowed to stir for about 1 hour at room temperature. Thereaction mixture was then concentrated on a rotary evaporator. Theresulting solid was dissolved in water, filtered and concentrated on therotary evaporator. The sample was freeze dried under vacuum and a solid170 (70%) was collected

Example 75 Alternative Synthetic Route to Tris(amino) 1,3,5-TriazineCompounds

[0839] The following reaction scheme represents a proposed andalternative synthetic routes to 1,3,5-triazines.

[0840] This scheme represents a modification of the synthetic routedescribed in the patent to prepare the tris-amino substituted1,3,5-triazines. Alternative leaving groups, X, could be used ascompared to cyanuric chloride (X═Cl) in the S_(N)Ar reaction with asequential addition of a nucleophilic amine in the presence of an acid(proton) scavenger to afford the tris-substituted 1,3,5-triazine withthe desired combination of amino groups.

Example 76 Alternative Synthetic Route to Tris(amino) 1,3,5-TriazineCompounds

[0841] The following reaction scheme represents a proposed andalternative synthetic routes to 1,3,5-triazines.

[0842] This scheme represents a modification of the synthetic routedescribed in the patent text to prepare the tris-amino substituted1,3,5-triazines. Bases, including excess amine reagent R₂NH, could beused as acid (proton) scavengers alternatively to the Huinig's base(iPr₂NEt) used routinely in our procedure. These bases can include otherorganic tertiary amine bases or ionic, inorganic bases. One can usestrong bases (NaH, KH, or RLi) to first deprotonate the amino monomerbefore addition to the cyanuric-X substrate. Additionally, one can use asolid supported base (e.g., resin-NR₂, a modified Hünig's base) as aproton scavenger. This potentially enables an easier isolation procedureand cleaner reaction products. Logically, one would use the appropriatesolvent or combination of solvents that is compatible with the base ofchoice for this procedure.

Example 77 Alternative Synthetic Route to Tris(amino) 1,3,5-TriazineCompounds

[0843] The following reaction scheme represents a proposed andalternative synthetic routes to 1,3,5-triazines.

[0844] This scheme represents a modification of the synthetic routedescribed in the patent to prepare the tris-amino substituted1,3,5-triazines. Using melamine as the starting material, the methodoutlined would involve three sequential reductive amination procedures.With control of addition, temperature, and pH, the choice of aldehydesor ketone, one can prepare tris-amino substituted triazines with thedesired combination of amino groups.

Example 78 Alternative Synthetic Route to Tris(amino) 1,3,5-TriazineCompounds

[0845] The following reaction scheme represents a proposed andalternative synthetic routes to 1,3,5-triazines.

[0846] This scheme represents a solid phase synthetic approach topreparing symmetrically or asymmetrically substituted tris-aminosubstituted 1,3,5-triazines. The resin should possess a readilycleavable linker group (L) and a leaving group (G) for attachment of anamino group. The scheme outlines the synthesis by initially attaching asimple amino group, NH₂, by reacting the resin with ammonia. Usingstandard, S_(N)Ar chemistry for substitution of a perhalogenated1,3,5-triazine, the triazine can be attached to the aminated resin.Sequential substitutions of the halogens on the triazine core withfunctionalized amines in the presence of an acid scavenger will producethe desired di-amino substituted 1,3,5-triazine. Cleavage of thetriazine from the resin tether will afford the tris-amino substitutedtriazine product. The free NH₂ moiety of the triazine can be furtheralkylated or functionalized using standard chemistry such reductiveamination or N-alkylation to give a completely functionalized tris-aminosubstituted 1,3,5-triazine.

Example 79 Alternative Synthetic Route to Tris(amino) 1,3,5-TriazineCompounds

[0847] The following reaction schemes (Schemes A and B) representproposed and alternative synthetic routes to 1,3,5-triazines.

[0848] These schemes represent variations on using the Suzuki couplingto synthesize tris-amino substituted 1,3,5-triazines. As illustrated inScheme A, one can sequentially react the amino groups of melamine withan alkyl or aryl boronic acid derivative in the presence of theappropriate palladium catalyst, additives and solvent to afford thesymmetric or asymmetric tris-amino substituted 1,3,5-triazines similarto previously described examples. In Scheme B, a tris-boronic acid1,3,5-triazine can be prepared from cyanuric chloride or bromide. Thisderivative can then be coupled with an aryl or alkyl amine, asillustrated in previous amine monomer descriptions, in the presence ofthe appropriate metal catalysts (e.g., Cu or Pd catalyst), additives andsolvent to afford the symmetric or asymmetric tris-amino substituted1,3,5-triazines.

Example 80 Proteoglycan Induction

[0849] Smooth muscle cells reach quiescence during serum starvationresulting in a blockade of DNA synthesis. To demonstrate the role ofperlecan (proteoglycan example) in SMC quiescence, cells were starved byremoving serum from the media. The cells used in this Example and theother examples herein were human aortic SMC, grown in basal mediumsupplemented with growth factors, bFGF and epidermal growth factor (EGF)(Clonetics, San Diego, Calif.).

[0850] SMC secretion of total PGs (proteoglycans) as well as perlecanwere determined in the presence or absence of one or more compounds ofthe present invention. PGs were radiolabeled with (³⁵S)sulfate byincubating the cells with (³⁵S)sulfate for 2 to 6 hours. Media PGs werecollected and purified by DEAE-cellulose chromatography. Cell-associatedPGs were assessed by extracting cells with 50 mM Tris buffer pH 7.4containing 4 M urea, 1% Triton X-100, 0.1 mM EDTA and 1 mM PMSF. Aqueoussolutions of (³⁵S)sulfate and (³H)leucine were from Amersham. Controlcells have no added compounds whereas treated cells have one or morecompounds of the present invention added.

[0851] To determine changes in PG levels, DEAE-cellulose chromatographywas performed. A DEAE-cellulose column was equilibrated with 50 mM Trisbuffer pH 7.4 containing 4 M urea, 0.1 M NaCl, 0.1 mM EDTA, 1 mM PMSFand 1% 3[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS).The column was washed with the same buffer and buffer containing 0.25 MNaCl and PG were eluted with the same buffer containing 0.5 M NaCl.Fractions containing radioactivity (³⁵SO₄) were pooled and dialyzedagainst MEM overnight and counted.

[0852] To determine the relative proportion of HSPG and chondroitinsulfate/dermatan sulfate proteoglycan (CS/DS PG), an aliquot of thepooled fraction was incubated in 50 mM sodium acetate buffer pH 5.2 with1 unit/ml each of heparanase and heparitinase or with 0.5 units ofchondroitin ABC lyase for 16 h at 37° C. Chondroitan ABC refers todifferent isomeric types of chondroitin, e.g. chondroitin A, chondroitinB, and chondroitin C. The reaction mixture was precipitated either with0.5 volumes of 1% cetyl pyridinium chloride or with 3 volumes of ethanolto precipitate undigested glycosaminoglycans. Radioactivity in thesupernatant and pellet was determined.

[0853] To determine changes in perlecan protein in response to thepresence of a compound, cells were grown in serum-free orserum-containing media in the presence of (³H)leucine for 24 h (steadystate). Cells were plated at low density (8×10⁴/well in 48 well plate,30-40% confluency) and cultured for 24 h (hour). Wells were thenreplenished with fresh medium containing no serum or 10% fetal bovineserum (FBS). Following another 24 h incubation, cells were labeled with(³H)thymidine for 6 h and radioactivity incorporated into the DNA wasdetermined by trichloroacetic acid (TCA) precipitation of the celllysate. (³H)thymidine was from NEN. Purified PG (0.5 M eluate) wereimmunoprecipitated by incubation with an anti-perlecan antibody(100-fold diluted) followed by precipitation with Protein A-Sepharose.Immunoprecipitates were analyzed by 5% SDS-PAGE. Perlecan (Mr>550 kDa)was identified by autoradiography. Control cells have no addedcompound(s) whereas treated cells have one or more compounds of thepresent invention added.

Example 81 Inhibition of Smooth Muscle Cell Proliferation

[0854] Purified perlecan from SMC medium by DEAE-cellulosechromatography was obtained using methods in Example I, and was testedfor its antiproliferative effects on SMC.

[0855] The addition of perlecan to serum-containing medium inhibited SMCgrowth by 70%. Sub-confluent SMC (40-50% confluence) were incubated inserum-free medium or 10% serum-containing medium with or withoutpurified perlecan for 24 h. DNA synthesis was then determined byincubating cells for another 5 h in medium containing (³H)thymidine. TCAprecipitable (DNA) thymidine counts were determined and expressed aspercentage of DNA synthesis in cells grown in 10% FBS.

[0856] This assay can be used to show the effect of a compound onperlecan directly by incubating the compound to perlecan first, thenperforming the assay. Alternatively, the cells can be pretreated with atleast one compound of the present invention to show indirect effects.Control cells have no added compounds whereas treated cells have one ormore compounds of the present invention added.

Example 82 Triazine Compounds in Smooth Muscle Cell Proliferation Assay

[0857] Human aortic smooth muscle cells (Clonetics) were used. Cellswere grown in basal medium containing 5% fetal bovine serum supplementedwith growth factors, basic fibroblast growth factor, epidermal growthfactor and insulin. To determine the effects triazine compounds of thepresent invention had on SMC proliferation, cells were plated at lowdensity (4000 cell per well in a 96 well plate) and cultured for 24 h.The cells were then serum starved for 24 h to induce quiescence. Freshgrowth medium containing no compound or 10 μM compound was then addedand further incubated for 24 h. Cell number was determined by using acell proliferation assay kit (Celltiter96 AQ_(ueous) from Promega).

[0858] The effects of different triazine compounds on smooth muscle cellproliferation are shown in FIG. 53. Many of the triazine compoundsinhibited SMC proliferation by greater than 70%.

Example 83 Induction and Measurement of Endothelial Heparanase Protein

[0859] Experiments were carried out on human microvascular endothelialcells (HMVEC) grown in 48-well plates (˜90% confluency). To induceheparanase activity, culture media was replaced with 200 μl Dulbecco'sModified Eagle's medium (DMEM) complemented with 1% bovine serum albumin(BSA) and with or without stimulants (5 ng/ml TGF-alpha, 1 ng/ml IL 1alpha, 200 ng/ml VEGF or other stimulants, cytokines, or inducers asrequired). The secreted proteins were analyzed by SDS/PAGE andheparanase protein was detected by immunoblotting using polyclonalanti-human heparanase antibody. The changes of heparanase expressiondetermined by densitometric analysis. The induction and measurement ofendothelial heparanase protein reported in the Tables herein werecarried out according to this Example.

Example 84 Preparation of Biotinylated HS

[0860] Heparan sulfate (HS) was biotinylated using biotin with extendedspacer arms using succinimidyl-6-(biotinamido) hexanoate (NHS-LC-Biotin)obtained from Pierce. About 0.5 ml HS solution (2 mg/ml in NaHCO3, pH8.5) was mixed with 0.05 ml of a freshly prepared solution ofNHS-LC-Biotin in dimethyl sulfoxide. The mixture was incubated at roomtemperature for 1 hour. Unconjugated biotin was removed bycentrifugation (10,000 RPM) through Microcon-3 filter (Millipore)followed by dilution with phosphate buffered saline (PBS). Thisprocedure was repeated five times to ensure complete removal of freebiotin. Unwanted aldehydes in the reaction were then quenched byincubation with one milliliter of Tris-glycine buffer (25 mM-183 mM, pH8.3) at room temperature for 20 minutes. The mixture was subjected tothree rounds of microfiltration as described above. Biotinylated HS (5mg/ml in PBS) was aliquoted and stored at −20° C. To obtain maximumbiotinylation, a 25-fold molar excess of biotin was used. Using HABAreagent, it was determined that the ratio of HS to biotin was 1:2.

[0861] The extent of biotinylation of HS was determined usingAvidin-HABA (Pierce Chemical Co). The HABA assay can be used over a widerange of pH and salt concentrations. HABA(4-hydroxyazobenzene-2′-carboxylic acid) is a dye that binds to avidinand can serve as an indicator of unoccupied binding sites. Avidincombines stoichiometrically with biotin, making it possible to use anyphysiochemical differences between avidin and the avidin-biotin complexas the basis of a qualitative and quantitative assay method for eithercomponent.

[0862] When HABA binds to avidin, there is a large spectral change inthe HABA dye. A new absorption band appears at 500 nm, which ischaracteristic of the quinoid form of the dye. The avidin-biotin complexdoes not bind HABA and because the dissociation constant of the complexis so low, the dye is stoichiometrically displaced by biotin.Consequently, the HABA assay can be the basis of both colorimetric andtitrimetric assays. The amount of avidin can be calculated directly fromthe increased absorbance at 500 nm, or the dye may be used as anindicator in a spectrophotometric titration with biotin.

[0863] The absorption band that results from the avidin-HABA complexdecreases proportionately when biotin is added. Since biotin has such ahigh affinity for avidin, it displaces the HABA dye. The unknown amountof biotin can be determined by preparing a standard curve using knownamounts of biotin to displace the HABA which bound to avidin, andplotting against the absorbance at 500 mu.

[0864] HABA solution was prepared by adding 24.2 mg of HABA (Pierce) to9.9 ml H20, and then adding 0.1 ml 1 M NaOH. Avidin-HABA reagent wasprepared by adding 10 mg of avidin and 600 gl of HABA solution to 19.4ml of phosphate buffered saline. To 1 ml of Avidin-HABA reagent in acuvette, 100 μI of biotinylated HS was added, and the optical densitywas measured at 500 nm in a spectrophotometer. A standard curve wasdetermined using known amounts of HABA. The decrease in optical densityof the HABA following the addition of biotinylated HS was determined.

Example 85 Heparanase Assay

[0865] Biotin-labeled HS made as described above was digested withheparanase, under both control and treated conditions, and the reactioncontaining undegraded and degraded HS was bound to in a biotin-bindingplate. Streptavidin, conjugated with an enzyme, was added to the bindingplate. Quantitation of the color reaction measured the amount ofavailable biotin binding sites. A decrease in color from a known amountreflects HS digestion by heparanase. Control conditions have no addedcompound of the present invention, and treated conditions have compoundsof the present invention added.

[0866] A lyophilized powder of heparanase (heparanase III obtained fromSeikagaku) containing 0.1 units of enzymatic activity was hydrated in100 μl of Reaction Buffer (3.33 mM calcium acetate pH 7.0, containing0.1 mg/ml BSA). This solution was then diluted to a workingconcentration of heparanase solution (0.01 micro-units to 1 milli-unit)in Reaction Buffer. Enzyme activity was defined by the manufacturer ofthe heparanase (Seikagaku) as follows: one unit of enzyme activity isdefined as amount required to generate 1 micromole of hexuronic acid perminute. Biotin-HS was diluted to a desired concentration in ReactionBuffer.

[0867] To determine heparanase activity, 10 μl of heparanase solution,with or without at least one of the compounds of the present invention,was mixed with 200 μl of the biotin-HS substrate in a 96 well plate. Thereaction was incubated at 43° C. for 1 hour. One hundred microliters ofthe reaction mixture was added to a hydrated biotin-binding plate(Chemicon) and incubated at 37° C. for 30 minutes. The biotin-bindingplates were hydrated with 200 μl of 1× Assay Buffer (Chemicon). Wellswere washed five times with 1× Assay Buffer and incubated with 100 μl of1:3000 diluted Streptavidin-Enzyme Conjugate (Chemicon) for 30 minutesat 37° C. The wells were washed five times with 1× Assay Buffer andincubated for 20 minutes with 100 μl of Substrate Solution (Chemicon).Color development in the wells was assessed by measuring the opticaldensity at 450 nm in a microplate reader (Labsystems, Muliskan Ascentmodel). Differences between the control and the treated conditionsindicate the heparanase modulating activity of the added compound orcompounds.

Example 86 AGE-Induced Inflammatory Response Determined by IL-6 ELISA

[0868] Human aortic endothelial cells (HAEC, Clonetics) were culturedaccording to manufacturer in growth medium (Clonetics): basal mediumcontaining human epidermal growth factor, hydrocortisone, vascularendothelial growth factor, heparin binding growth factor-B, longR3-insulin-like growth factor-1, ascorbic acid, gentamicin/amphotericinand 5% FBS. These cell were allowed to reach at least 90% confluencybefore subjected to experimental treatments. Glycated human serumalbumin (G-HSA) was from US Biologicals. Tumor necrosis factor α wasfrom R&D Systems.

[0869] Endothelial cells were treated with control medium or mediumcontaining 10 to 100 ng/ml TNF-α or 300 μg/ml glycated-HAS (treatedcells or treatments) for 24 hrs, in control and compound-addedduplicates, containing 10 μM compound. All treatments, compound-addedand controls were carried out in serum free media containing 0.2%albumin. Media from all conditions were collected and used for IL-6ELISA.

[0870] IL-6 ELISA was carried out using human IL-6 DuoSet ELISAdevelopment kit as described by manufacturer (R&D Systems). Mouseanti-human II-6 was used as the capture antibody (2 ug/ml) andbiotinylated goat anti-human IL-6 (200 ng/ml) was used as the detectionantibody. Culture media were incubated with capture antibody (in 96well) for 2 h at room temperature. Wells were washed three times withwash buffer (0.05% tween-20 in phosphate buffered saline (PBS) pH 7.4)followed by incubation with detection antibody for 2 h at roomtemperature. Following three washes wells were incubated withStreptavidin-HRP for 20 min. Color development was read at 450 nm in aMicroplate reader.

[0871] The effects of compounds of the present invention on G-HSAinduced IL-6 are shown in FIG. 54. G is G-HSA, and C is control, notreatment with compounds or G-HSA. Endothelial cells under basalconditions secreted about 25 pg/ml of IL-6. Incubation of endothelialcells with G-HSA induced a 3 fold increase in IL-6 secretion byendothelial cells. Addition of compounds of the present invention, asindicated by each compound's number, to G-HSA containing mediasignificantly reduced endothelial secretion of IL-6. These inhibitoryeffects varied, the most effective compounds showed an 80% decrease inIL-6 secretion. These data show the compounds of the present inventionhave anti-inflammatory activity.

Example 87 Cytotoxicity/Lactate Dehydrogenase Assay

[0872] An appropriate number of cells are plated in four 96-well plates,one plate for “day 0” and three plates for days 1-3. Cells are treatedwith at least one compound of the present invention in varyingconcentrations with and without the apoptosis inducer cisplatin (2 μM)(“+cis” or “−cis”). Unteated cells are also assayed with and withoutcisplatin. After transfection, the plates are incubated at 37° C.overnight.

[0873] An appropriate number of cells are plated in four 96-well plates,one plate for “day 0” and three plates for days 1-3. Cells are treatedwith at least one compound of the present invention in varyingconcentrations. The negative control cells have normal media conditions,a duplicate set of wells is treated with the composition in which thecompound is provided, but there is no added compound and the positivecontrol cells are treated with the apoptosis inducer cisplatin (2 μM).All of the cells are transfected with a vector having a promoter that isresponsive to apoptosis conditions. When apoptosis occurs, the promoteris turned on and the lactic dehydrogenase gene is activated and theenzyme protein is made and active. Activity is easily detected with acolor change. After transfection, the plates are incubated at 37° C.overnight.

[0874] About 8 mls of warmed alpha MEM LDH lysis buffer (2% Triton ×100)and about 8 mls of culture media (½ dilution) are combined. Two 96-wellv-bottom plates are prepared, one labeled “lysis” and one labeled“supernatant.” To lyse the cells, about 200 μl Alpha MEM lysis buffer(diluted ½) is added to one test plate from which the supernatant hasbeen removed and added to the plate labeled supernatant. After mixing,about 200 μl of lysed cells are transferred to the lysis plate. Both thelysis and supernatant plates are centrifuged at about 1600 rpm for about10 min. After centrifugation, about 100 μl of both the supernatant orlysate is transferred to corresponding 96-well flat-bottomed plates.

[0875] The assay for cytotoxicity uses the Cytotoxicity Detection Kit(LDH) from Roche Diagnostics Corp. (Indianapolis, Ind.). Using thedirectionts provided, the dye solution is mixed and added each well ofthe lysate and supernatant plate and incubated for up to 20-25 min at15-25° C. in the dark.

[0876] The difference in the amount of lactate dehydrogenase releasedfrom cells in untreated cells when compared to cells treated withcisplatin or compounds of the present invention having cytoxic activityshows the cytotoxic activity of the compounds tested.

Example 88 Synthesis of 4-Benzyloxy-3-chloro-phenylamine (E1)

[0877]

[0878] To 4-amino-2-chlorophenol (7.23 g, 50 mmol) dissolved in acetone(250 mL), was added potassium carbonate (6.94 g, 50 mmol), followed bythe addition of benzyl chloride (5.8 mL, 50 mmol), tetrabutylammoniumbromide (TBAB) (1.66 g, 5 mmol), and potassium hydroxide (2.84 g, 50mmol). The reaction mixture was allowed to stir at reflux overnightunder nitrogen. The reaction mixture was extracted three times usingdichloromethane; the combined organic layers were washed with brine anddried over potassium carbonate. The sample was concentrated and theresulting oil was dried overnight under vacuum. The material waspurified by silica gel flash column chromatography eluting with 50:50v:v hexanes:ethyl acetate and the collected fractions were concentratedin vacuo to give E1 (9.3 g, 80%); mp 54° C.; HPLC: Inertsil ODS 3V C18,40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 14.8 min,98.7% purity; MS (TOF ES+) m/z 234 (M+H, 100).

Example 89 Synthesis ofN-(4-Benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(E2)

[0879]

[0880] To cyanuric chloride (3.148 g, 17.0 mmol) dissolved inacetonitrile (70 mL) stirring at −20° C., was added a solution of4-benzyloxy-3-chloro-phenylamine (4.0188 g, 17.0 mmol) in acetonitrile(40 mL) followed by the addition of diisopropylethylamine (DIEA) (3 mL,17.0 mmol), and was stirred at −20° C. for 1 hour under a nitrogenatmosphere. The mixture was allowed to warm to room temperature and thencycloheptylamine (2.2 mL, 17.0 mmol) was added in anhydrous acetonitrile(5 mL) followed by addition of a DIEA (3.2 mL, 18.7 mmol), and themixture was stirred at room temperature overnight. To this reactionmixture was added DIEA (3.2 mL, 18.7 mmol) followed by addition ofN-methyl-4-(methylamino)piperidine (2.5 mL, 17.0 mmol) and was stirredand heated at reflux overnight. The reaction mixture was extracted 3times with methylene chloride; the combined organic layers were washedone time with brine and dried over anhydrous potassium carbonate. Theorganic layer was concentrated in vacuo and dried overnight undervacuum. Column chromatography (90:9:1 v:v:vdichloromethane:methanol:ammonium hydroxide) yielded E2 (619 mg, 7%); mp84° C.; HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 37.1 min, 99.4% purity; MS (TOF ES+)m/z 550 (M+H, 100), 276 (M+2H, 34.9), 275.5 (83.9).

Example 90 Synthesis ofN-(4-benzyloxy-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine(E3)

[0881] Compound E3 was obtained as a side product (2.63 g) via columnchromatography (silica gel; 90:9:1 v:v:vdichloromethane:methanol:ammonium hydroxide); mp 74° C.; HPLC: InertsilODS 3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm,R_(t) 38.6 min, 99.8% purity; MS (TOF ES+) m/z 536 (M+H, 100), 269(44.6), 268.5 (M+2H, 97.9).

Example 91 Synthesis of4-[4-Cycloheptylamino-6-(methyl-piperidin-4-yl-amino)-[1,3,5]triazin-2-ylaminol-phenol(E4)

[0882]

[0883] In a dry round bottomed flask was added 10% Pd/C (301.5 mg) andwas wetted with 3-5 drops of water. Nitrogen was blown over the Pd/C forabout 5 minutes, then methanol (10 mL) was added cautiously and nitrogenwas blown on the mixture for another 5 minutes. Triazine E3 (251.8 mg,0.50 mmol) dissolved in methanol (5 mL) was added followed by ammoniumformate (381.5 mg, 6.0 mmol), and the reaction was stirred and heatedreflux for about 1.5 hours. Methylene chloride was added and allowed tocool to room temperature. The mixture was filtered by vacuum throughCelite with a methylene chloride rinsing and the filtrate wasconcentrated in vacuo. The material was purified by silica gel flashchromatography eluting with 90:9:1 by volume methylenechloride:methanol:ammonium hydroxide, and the collected fractions weredried over anhydrous magnesium sulfate, filtered and then concentratedin vacuo to give a light yellow solid (E4) (98 mg, 44%); mp 130° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄(0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 3.2 min, 99.6% purity; MS (TOF ES+) m/z 412(M+H, 100), 235.6 (87.2), 206.6 (16.1).

Example 92 Synthesis of4-{4-Cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-1,3,5]triazin-2-ylamino}-phenol(E5)

[0884]

[0885] In a dry flask was added 10% Pd/C (301.0 mg) which was wettedwith 3-5 drops of water. Nitrogen was blown over the Pd/C for about 5minutes, then methanol (10 mL) was added cautiously, and nitrogen wasblown over the mixture for another 5 minutes. Triazine E2 (250.5 mg,0.45 mmol) dissolved in methanol (5 mL) was added followed by ammoniumformate (340.5 mg, 5.4 mmol), and the reaction was stirred and heated atreflux for about 1.5 hours. Methylene chloride was added and allowed tocool to room temperature. The mixture was filtered by vacuum throughCelite with a methylene chloride rinsing and the filtrate wasconcentrated in vacuo. The crude material was dried overnight undervacuum to give a light brown solid (E5) (144 mg, 70%); mp 157° C.; HPLC:Inertsil ODS 3V C18, 40:30:30 v:v:v [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 3.0 min, 93.8% purity; MS (TOF ES+) m/z426.3 (M+H, 20.9), 234.1 (100).

Example 93 Synthesis of2-Chloro-4-(4,6-dichloro-[1,3,5]triazin-2-ylamino)-phenol (E6)

[0886]

[0887] Cyanuric chloride (12.95 g, 70.0 mmol) was dissolved in acetone(200 mL), the solution was cooled to 0-5° C. with an ice bath, and4-amino-2-chlorophenol (10.09 g, 70.0 mmol) in acetone (100 mL) wasadded. The mixture was stirred at 0-5° C. for 1 hour under nitrogen. Thereaction mixture was extracted three times using dichloromethane; thecombined organic layer were washed with brine and dried over sodiumsulfate. The sample was concentrated in vacuo, and the resulting solidwas dried overnight under vacuum to yield E6 (20.2 g, 99%); mp 180° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 v:v:v [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 11.6 min, 99.4% purity; MS (TOF ES+)miz 293 (97.5), 291 (M+H, 100).

Example 94 Synthesis of2-Chloro-4-(4-chloro-6-cycloheptylamino-[1,3,5]triazin-2-ylamino)-phenol(E7)

[0888]

[0889] To E7 (1.06 g, 3.4 mmol) dissolved in acetone (25 mL) was added asolution of cycloheptylamine (0.44 mL, 3.4 mmol) in acetone (5 mL)followed by addition of 2.5 N NaOH (1.4 mL, 3.4 mmol) and water (3.5mL). The reaction mixture stirred and heated at reflux for 3 hours. Thereaction mixture was extracted three times using dichloromethane; thecombined organic layers were washed with brine and dried over sodiumsulfate. The sample was concentrated and was dried overnight undervacuum to yield a light brown solid (E7) (1.25 g, 99%); mp 91° C.; HPLC:Inertsil ODS 3V C18, 40:30:30 v:v:v [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 42.9 min, 92.1% purity; MS (TOF ES+)m/z 370 (65.9), 368 (M+H, 100).

Example 95 Alternative Synthesis of2-Chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino}-[1,3,5]triazin-2-ylamino)-phenol(138)

[0890] To E7 (1.00 g, 2.7 mmol) dissolved in THF (25 mL) was added asolution of N-methyl-4-(methylamino)-piperidine (0.45 mL, 3.1 mmol) inTHF (5 mL) followed by addition of 2.5 N NaOH (1.1 mL, 2.7 mmol) and 2.5mL of water. The reaction mixture was stirred and heated at refluxovernight. The reaction mixture was extracted three times usingdichloromethane, the combined organic layers were washed with a brineand dried over sodium sulfate. The sample was concentrated and theresulting solid was dried overnight under vacuum. Column chromatography(silica gel, 100% methanol) yielded an off-white solid (138) (177 mg,14%); mp 68° C.; HPLC: Inertsil ODS 3V C18, 40:30:30 v:v:v [KH₂PO₄ (0.01M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 4.7 min, 99.6% purity; MS (TOFES+) m/z 460 (M+H, 55.3), 251 (100), 224 (51.1).

Example 96 Synthesis ofN-(1-Benzyl-piperidin-4-yl)-N′-(3-fluoro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-diamine(E8)

[0891]

[0892] To 127 (6.00 g, 16.4 mmol) dissolved in THF (100 mL) was added4-amino-1-benzylpiperidine (3.6 mL, 18.9 mmol) in THF (5 mL), followedby addition of H₂O (17 mL) and 2.5 N NaOH (6.6 mL, 16.4 mmol). Thereaction mixture was stirred and heated at reflux for about 12 hoursunder nitrogen. The reaction mixture was extracted three times withdichloromethane; the combined organic layers were washed with brine anddried over potassium carbonate. The sample was filtered, concentrated,and the resulting solid was dried overnight under vacuum. Columnchromatography (93:6:1 v:v:v dichloromethane:methanol:ammoniumhydroxide) gave a light yellow solid (E8) (2.98 g, 35%); mp 87° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄(0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 11.1 min, 93.9% purity; MS (TOF ES+)m/z 520 (M+H, 66.3), 430 (100).

Example 97 Alternative Synthesis ofN-(1-Benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-diamine(134)

[0893]

[0894] To 133 (6.04 g, 15.7 mmol) dissolved in THF (100 mL) was added4-amino-1-benzylpiperidine (3.4 mL, 18 mmol) in THF (5 mL), followed byaddition of water (16 mL) and 2.5 N NaOH (6.3 mL, 15.7 mmol). Thereaction mixture was stirred and heated at reflux for about 12 hoursunder a nitrogen atmosphere. The reaction mixture was extracted threetimes with dichloromethane; the combined organic layers were washed withbrine and dried over potassium carbonate. The sample was filtered,concentrated, and the resulting solid was dried overnight under vacuum.Column chromatography (93:6:1 v:v:v dichloromethane:methanol:ammoniumhydroxide) gave a light yellow solid (134) (3.71 g, 44%). mp 90° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 14.6 min, 99.5% purity; MS (TOF ES+)m/z 538 (27.8), 536 (M+H, 72.1), 448 (39.2), 446 (100).

Example 98 Synthesis ofN-Cycloheptyl-N′-(4-methoxy-phenyl)-N″-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine(E9)

[0895]

[0896] In a dry round bottomed flask, 10% Pd/C (600.2 mg) was added andwas wetted with 3-5 drops of water. Nitrogen was blown over the Pd/C forabout 5 minutes, then methanol (25 mL) was added cautiously, andnitrogen was again blown over the mixture for about 5 minutes. Triazine134 (501.2 mg, 0.93 mmol) dissolved in methanol (15 mL) was addedfollowed by ammonium formate (708.4 mg, 11.2 mmol), and the reaction wasstirred and heated at reflux for about 1.5 hours. Methylene chloride wasadded and allowed to cool to room temperature. The mixture was filteredby vacuum through Celite with a methylene chloride rinsing, the filtratewas concentrated, and the material was dried overnight under vacuum.Column chromatography (90:9:1 v:v:v dichloromethane:methanol:ammoniumhydroxide) yielded a light yellow solid (E9) (226 mg, 55%); mp 118° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 4.6 min, 99.6% purity; MS (TOF ES+) m/z412 (M+H, 39.8), 247.9 (63), 227.3 (100).

Example 99 Synthesis of6-Chloro-N-cyclopropyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(E10)

[0897]

[0898] To 124 (2.01 g, 7.0 mmol) dissolved in acetone (40 mL) was addeda solution of cyclopropylamine (0.5 mL, 7 mmol) in acetone (5 mL)followed by addition of NaOH (2.8 mL, 2.5 N, 7.0 mmol) and 8 mL ofwater. The reaction was stirred and heated at reflux for 3 hours. Thereaction mixture was poured over crushed ice. The solid that formed wascollected by vacuum filtration and was dried overnight under vacuum toafford unpurified E10 (1.8 g). {A separate reaction [124, (2.00 g, 7mmol) and cyclopropylamine (0.5 mL, 7 mmol)] yielded unpurified E10 (1.9g).} The two lots (1.8 g and 1.9 g) were combined for purification.Column chromatography (50:50 v:v hexane:ethyl acetate) yielded a faintyellow solid (E10) (2.89 g, 67%); mp 186° C.; HPLC: Inertsil ODS 3V C18,40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 16.6 min,94.9% purity; MS (TOF ES+) m/z 310 (M+H, 100), 312 (44.4).

Example 100 Synthesis ofN-Cyclopropyl-N′-(3-flouro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(E11)

[0899] To E10 (1.04 g, 3.2 mmol) dissolved in THF (40 mL) was added asolution of N-methyl-4-(methyl-amino)-piperidine (0.5 mL, 3.2 mmol) inTHF (2 mL) followed by addition of NaOH (1.3 mL, 2.5 N, 3.2 mmol) and3.5 mL of water. The reaction mixture stirred and heated at reflux for 2hours. The reaction mixture was extracted 3 times using dichloromethane;the combined organic layers were washed with brine and dried overpotassium carbonate. The sample was filtered, concentrated, and theresulting solid was dried overnight under vacuum. Column chromatography(90:9:1 v:v:v dichloromethane:methanol:ammonium hydroxide) yielded alight yellow solid (E11) (164 mg, 13%); mp 94° C.; HPLC: Inertsil ODS 3VC18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 3.2min, 96.7% purity; MS (TOF ES+) m/z 402.1 (M+H, 100), 231 (41.5), 202.1(6).

Example 101 Synthesis ofN-Cyclopropyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine(E12)

[0900]

[0901] To E10 (1.05 g, 3.2 mmol) dissolved in THF (40 mL) was added asolution of 2-(aminomethyl)-1-ethylpyrrolidine (0.5 mL, 3.2 mmol) in THF(2 mL) followed by addition of NaOH (1.3 mL, 2.5 N, 3.2 mmol) and 3.5 mLof water. The reaction mixture was stirred and heated at reflux for 2hours. The reaction mixture was extracted 3 times using dichloromethane;combined organic layers were washed with brine and dried over potassiumcarbonate. The sample was filtered, concentrated, and dried overnightunder vacuum. Column chromatography (90:9:1 v:v:vdichloromethane:methanol:ammonium hydroxide) yielded a light yellowsolid (E12) (755 mg, 59%); mp 68° C.; HPLC: Inertsil ODS 3V C18,40:30:30 [KH₂PO₄ (0.01 M, pII 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 3.2 min,95.5% purity; MS (TOF ES+) m/z 402.1 (M+H, 100), 231. (23.4).

Example 102 Synthesis of6-Chloro-N-(3-chloro-4-methoxy-phenyl)-N′-cyclopropyl-[1,3,5]triazine-2,4-diamine(E13)

[0902]

[0903] To 101 (2.01 g, 6.5 mmol) dissolved in acetone (35 mL) was addeda solution of cyclopropylamine (0.45 mL, 6.5 mmol) in acetone (5 mL)followed by addition of NaOH (2.6 mL, 2.5 N, 7.0 mmol) and 6.5 mL ofwater. The reaction mixture was stirred and heated at reflux for 3hours. The reaction mixture was extracted 3 times with dichloromethane;the combined organic layers were washed with brine and dried overpotassium carbonate. The sample was filtered, concentrated, and driedovernight under vacuum. Column chromatography (50:50 v:v hexane:ethylacetate) yielded a faint yellow solid (E13) (1.12 g, 53%); mp 172° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 24.6 min, 99% purity; MS (TOF ES+) m/z328 (72.6), 326 (M+H, 100).

Example 103 Synthesis ofN-Cyclopropyl-N′-(3-chloro-4-methoxy-phenyl)-N″-methyl-“N-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine(E14)

[0904] To E13 (0.88 g, 2.7 mmol) dissolved in THF (40 mL) was added asolution of N-methyl-4-(methyl-amino)-piperidine (0.4 mL, 2.7 mmol) inTHF (2 mL) followed by addition of NaOH (1.1 mL, 2.5 N, 2.7 mmol) and 3mL of water. The reaction mixture was stirred and heated at reflux for 2hours. The reaction mixture was extracted 3 times with dichloromethane;the combined organic layers were washed with brine and dried overpotassium carbonate. The sample was filtered, concentrated, and theresulting solid was dried overnight under vacuum. Column chromatography(90:9:1 v:v:v dichloromethane:methanol:ammonium hydroxide) yielded alight yellow solid (E14) (93 mg, 8.3%); mp 92° C.; HPLC: Inertsil ODS 3VC18, 40:30:30 [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 3.4min, 96.9% purity; MS (TOF ES+) m/z 418.1 (M+H, 100), 210.1 (12.3).

Example 104 Synthesis of6-Chloro-N,N′-bis-(3fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine(E15)

[0905]

[0906] To 124 (1.09 g, 3.5 mmol) dissolved in acetone (25 mL) was added3-fluoro-p-anisidine (0.54 g, 3.5 mmol) in acetone (5 mL) followed byaddition of 2.5 N NaOH (1.4 mL, 3.5 mmol) and water (3.5 mL). Thereaction mixture was stirred and heated at reflux for 3 hours. Thereaction mixture was extracted three times using dichloromethane; thecombined organic layers were washed with brine and dried over potassiumcarbonate. The sample was filtered, concentrated in vacuo, and theresulting solid was dried overnight under vacuum to yield white solid(E15) (1.332 g, 97%); mp 194° C.; HPLC: Inertsil ODS 3V C18, 40:30:30v:v:v [KH₂PO₄ (0.01 M, pH 3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 33.2 min,97.6% purity; MS (TOF ES+) m/z 396 (35.0); 394 (M+H, 100).

Example 105 Synthesis ofN-(1-Ethyl-pyrrolidin-2-ylmethyl)-N′,N″-bis-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine(E16)

[0907] To E15 (1.01 g, 2.5 mmol) dissolved in THF (25 mL) was added asolution of 2-(aminomethyl)-ethyl pyrrolidine (0.42 mL, 2.9 mmol) in THF(5 mL) followed by addition of 2.5 N NaOH (1.0 mL, 2.5 mmol) and 2.5 mLof water. The reaction mixture was stirred and heated at refluxovernight. The reaction mixture was extracted three times usingdichloromethane; the combined organic layers were washed with brine anddried over potassium carbonate. The sample was filtered, concentrated invacuo, and the resulting solid was dried overnight under vacuum. Columnchromatography (90:9:1 v:v:v dichloromethane:methanol:ammoniumhydroxide) yielded a light yellow solid (E16) (570 mg, 47%); mp 66° C.;HPLC: Inertsil ODS 3V C18, 40:30:30 v:v:v [KH₂PO₄ (0.01 M, pH3.2):CH₃OH:CH₃CN], 264 nm, R_(t) 4.7 min, 99.3% purity; MS (TOF ES+) m/z486 (68.4, M+H), 264 (100), 244 [(M+2H)++, 48.1).

Example 106 Synthesis of 1-[4-(3-chloro-4-methoxyanilino)-cycloheptylamino-1,3,5-triazin-2-yl]-2-azoloamymethanol (E17)

[0908]

[0909] To 133 (0.2 g, 0.52 mmol) in Dowtherm (5 mL) was added slowlyprolinol (0.16 g, 1.57 mmol) and the mixture was heated to 150-160° C.for 5 hours with stirring. After completion of the reaction the mixturewas cooled to 25° C. and then transferred directly to a silica gelcolumn for purification purpose. The column was eluted using 1:1 (byvolume) EtOAc-Petroleum ether to afford E17 as a light brown solid (0.14g, 60%), mp 82-84° C.; HPLC: Inertsil ODS 3V (250×4.6 mm) 5 micron 50:50[0.01 M KH₂PO₄ (0.1% TFA):CH₃CN], 223 nm, R_(t) 11.86 min, 97.49%purity); MS (CI): m/z 447 (M⁺, 100), 416 (35).

Example 107 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(4-methylpiperzino)-1,3,5-triazine-4,2-diamine(E18)

[0910]

[0911] To a solution of 133 (0.5 g, 1.31 mmol) in 1,4-dioxane (5 mL) wasadded slowly a solution of 1-methylpiperizine (0.13 g, 1.31 mmol) in1,4-dioxane (5 mL) followed by 2.5 N sodium hydroxide solution (0.5 mL,1.31 mmol) and water (1.2 mL). The mixture was heated to reflux for2.5-3 hours with stirring under nitrogen atmosphere and then cooled to25° C. and diluted with water (10 mL). The mixture was then stirred for10-15 min., the precipitated solid was filtered off and dried undervacuum to afford the title compound E18 as an off-white solid (0.5 g,86%). mp 120-122° C.; HPLC: Inertsil ODS 3V (250×4.6 mm) 5 microns 65:35[0.01 M KH₂PO₄ (0.1% TFA):CH₃CN], 223 nm, R_(t) 10.23 min, 98.50%purity); MS (CI): m/z 446 (M+H, 100), 375 (25).

Example 108 Synthesis of3-[4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazin-2-yloxy]-2-ethyl-4H-4-pyranone(E19)

[0912]

[0913] A mixture of 133 (0.3 g, 0.78 mmol),2-ethyl-3-hydroxy-4H-pyran-4-one (0.11 g, 0.78 mmol) and K₂CO₃ (0.54 g,3.91 mmol) in dimethylformamide (15 mL) was stirred at 80° C. for 6hours under nitrogen atmosphere. After completion of the reaction themixture was cooled to 25° C. and diluted with water (50 mL). Theprecipitated solid was filtered off and dried under vacuum to afford thetitle compound E19 as an off-white solid (0.2 g, 52%). mp 124-126° C.;HPLC: Inertsil ODS 3V (250×4.6 mm) 5 microns 50:50 [0.01 M KH₂PO₄ (0.1%TFA):CH₃CN], 223 nm, R_(t) 18.47 min, 98.18% purity); MS (CI): m/z 486(M⁺, 100), 382 (90).

Example 109 Synthesis of1-[3-{4-(3-chloro-4-methoxyanilino)-6-cycloheptylamino-1,3,5-triazine-2-yloxy}piperidino]-1-ethanone(E20)

[0914]

[0915] A mixture of N-acetyl-3-hydroxypiperidine (0.335 g, 2.27 mmol)and sodium hydroxide (95 mg, 2.27 mmol) in benzene (10 mL) was heated toreflux for 2 hours with stirring under nitrogen atmosphere and thencooled to 25° C. followed by the addition of compound 133 (0.3 g, 0.78mmol) at same temperature. The mixture was heated to reflux for 6 hours,concentrated under vacuum and diluted with water (10 mL). Theprecipitated solid was filtered off and purified by columnchromatography to afford the title compound E20 as an off-white solid(0.35 g, 91%). mp 138-140° C.; HPLC: Inertsil ODS 3V (250×4.6 mm) 5microns [solvent A=0.01 M KH₂PO₄ (pH 7.0); solvent B=CH₃CN], Gradientelution program: T/% B=0/60, 10/60,25/80, 40/80, 45/60, 50/60; 268 nm,R_(t) 17.20 min, 91.70% purity; MS (CI): m/z 489 (M⁺, 100).

Example 110 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-isopropoxy-1,3,5-triazine-2,4-diamine(E21)

[0916]

[0917] A mixture of isopropanol (2 mL) and sodium hydroxide (95 mg, 2.27mmol) in benzene (10 mL) was heated to reflux for 2 hours with stirringunder nitrogen atmosphere and then cooled to 25° C. followed by theaddition of compound 133 (0.3 g, 0.78 mmol) at same temperature. Themixture was heated to reflux for 6 hours, concentrated under vacuum anddiluted with water (10 mL). The precipitated solid was filtered off andpurified by column chromatography (1-2% MeOH—CHCl₃) to afford the titlecompound E21 as an off-white solid (0.30 g, 94%). mp 136-138° C.; HPLC:Inertsil ODS 3V (250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄ (pH7.0); solvent B=CH₃CN], Gradient elution program: T/% B=0/60,10/60,25/80, 40/80, 45/60, 50/60; 268 nm, R_(t) 30.06 min, 98.78%purity; MS (CI): m/z 406 (M+H, 100).

Example 111 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine(E22)

[0918]

[0919] A mixture of prolinol (0.0243 g, 2.27 mmol) and sodium hydroxide(95 mg, 2.27 mmol) in benzene (10 mL) was heated to reflux for 2 hourswith stirring under nitrogen atmosphere and then cooled to 25° C.followed by the addition of compound 133 (0.3 g, 0.78 mmol) at sametemperature. The mixture was heated to reflux for 6 hours, concentratedunder vacuum and diluted with water (10 mL). The precipitated solid wasfiltered off and purified by column chromatography (1-2% MeOH—CHCl₃) toafford the title compound E22 as an off-white solid (0.35 g,quantitative). mp 84-86° C.; HPLC: Inertsil ODS 3V (250×4.6 mm) 5microns [solvent A=0.01 M KH₂PO₄ (pH 7.0); solvent B=CH₃CN], Gradientelution program: T/% B=0/60, 10/60,25/80, 40/80, 45/60, 50/60; 268 nm,R_(t) 24.67 min, 99.59% purity; MS (CI): m/z 447 (M+H, 100).

Example 112 Synthesis of1-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-3-ol](E23)

[0920]

[0921] A mixture of 3-hydroxypiperidine (0.198 g, 1.96 mmol) and sodiumhydroxide (79 mg, 1.96 mmol) in benzene (10 mL) was heated to reflux for2 hours with stirring under nitrogen atmosphere and then cooled to 25°C. followed by the addition of compound 133 (0.25 g, 0.65 mmol) at sametemperature. The mixture was heated to reflux for 6 hours, concentratedunder vacuum and diluted with water (10 mL). The precipitated solid wasfiltered off and purified by column chromatography (1-2% MeOH—CHCl₃) toafford the title compound E23 as an off-white solid (0.06 g, 21%). mp100-102° C.; HPLC: Hichrom RPB (250×4.6 mm) 5 microns [solvent A=0.01 MKH₂PO₄ (pH 5.5); solvent B=CH₃CN], Gradient elution program: T/% B=0/20,10/20,25/80, 40/80, 55/60, 65/20, 70/20; 270 nm, R_(t) 41.95 min, 98.43%purity; MS (CI): m/z 447 (M+H, 100).

Example 113 Synthesis of1-[4-(3-chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazine-2-yl]-piperidin-4-ol](E24)

[0922]

[0923] A mixture of 4-hydroxypiperidine (0.198 g, 1.96 mmol) and sodiumhydroxide (79 mg, 1.96 mmol) in benzene (10 mL) was heated to reflux for2 hours with stirring under nitrogen atmosphere and then cooled to 25°C. followed by the addition of compound 133 (0.25 g, 0.65 mmol) at sametemperature. The mixture was heated to reflux for 6 hours, concentratedunder vacuum and diluted with water (10 mL). The precipitated solid wasfiltered off and purified by column chromatography (1-2% MeOH—CHCl₃) toafford the title compound E24 as an off-white solid (0.20 g, 58%). mp142-144° C.; HPLC: Hichrom RPB (250×4.6 mm) 5 microns [solvent A=0.01 MKH₂PO₄ (pH 5.5); solvent B=CH₃CN], Gradient elution program: T/% B=0/20,10/20,25/80, 40/80, 55/60, 65/20, 70/20; 270 nm, R_(t) 40.56 min, 98.60%purity; MS (CI): m/z 447 (M+H, 100).

Example 114 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-2-azolanylmethoxy)-1,3,5-triazine-2,4-diamine(E25)

[0924]

[0925] A mixture of N-methyl prolinol (0.226 g, 1.96 mmol) and sodiumhydroxide (79 mg, 1.96 mmol) in benzene (10 mL) was heated to reflux for2 hours with stirring under nitrogen atmosphere and then cooled to 25°C. followed by the addition of compound 133 (0.25 g, 0.65 mmol) at sametemperature. The mixture was heated to reflux for 6 hours, concentratedunder vacuum and diluted with water (3 mL). The gum which separated wascollected (by decanting out the liquid portion) and purified by columnchromatography (1-3% MeOH—CHCl₃) to afford the title compound E25 as asemi-solid (0.06 g, 20%). HPLC: Symmetry shield RP18 (250×4.6 mm) 5microns [solvent A=0.01 M KH₂PO₄ (pH 3.0); solvent B=CH₃CN], Gradientelution program: T/% B=0/35, 10/35, 40/80, 50/80, 55/35, 60/35; 270 nm,R_(t) 15.51 min, 99.55% purity; MS (CI): m/z 461 (M+H, 100).

Example 115 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1-methyl-4-piperidyloxy)-1,3,5-triazine-2,4-diamine(E26)

[0926]

[0927] A mixture of N-methyl piperidinol (0.22 g, 1.96 mmol) and sodiumhydroxide (79 mg, 1.96 mmol) in benzene (10 mL) was heated to reflux for2 hours with stirring under nitrogen atmosphere and then cooled to 25°C. followed by the addition of compound 133 (0.25 g, 0.65 mmol) at sametemperature. The mixture was heated to reflux for 6 hours, concentratedunder vacuum and diluted with water (10 mL). The precipitated solid wasfiltered and purified by column chromatography (5-10% MeOH—CHC]₃) toafford the title compound E26 as a pale yellow solid (0.16 g, 53%).HPLC: Hichrom RPB (250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄ (pH7.0); solvent B=CH₃CN], Gradient elution program: T/% B=0/60, 10/60,25/80, 40/80, 45/60, 50/60; 270 nm, R_(t) 35.21 min, 98.09% purity; MS(CI): m/z 461 (M+H, 50), 364 (100).

Example 116 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(1,4-thiazinan-4-yl)-1,3,5-triazine-4,2-diamine(E27)

[0928]

[0929] A mixture of thiomorpholine (97 mg, 0.942 mmol), sodium hydroxide(31.4 mg, 0.785 mmol) and water (0.5 mL) in 1,4-dioxane (15 mL) washeated to reflux for 3 hours with stirring under nitrogen atmosphere andthen cooled to 25° C. followed by the addition of compound 133 (0.30 g,0.785 mmol) at same temperature. The mixture was heated to reflux for 12hours, concentrated under vacuum and diluted with water (20 mL). Themixture was then extracted with EtOAc (2×20 mL). The combined organiclayers were washed with water (2×10 mL), dried over anhydrous Na₂SO₄ andconcentrated under vacuum. The residue thus obtained was purified bycolumn chromatography (10-20% EtOAc-Hexane) to afford the title compoundE27 as a pale yellow solid (0.26 g, 74%). mp 78-80° C.; HPLC: InertsilODS 3V (250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄; solventB=CH₃CN], A:B=20:80; 220 nm, R_(t) 20.36 min, 97.15% purity; MS (CI):m/z 449 (M+H, 100).

Example 117 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(2-fluorophenoxy)-1,3,5-triazine-4,2-diamine(E28)

[0930]

[0931] A mixture of compound 133 (0.3 g, 0.785 mmol), 2-fluorophenol(0.106 g, 0.942 mmol) and potassium carbonate (541 mg, 3.93 mmol) indimethylformamide (3 mL) was heated to 80° C. for 12 hours with stirringunder nitrogen atmosphere. The mixture was then cooled, diluted withwater (20 mL) and extracted with EtOAc (2×20 mL). The combined organiclayers were washed with water (2×10 mL), dried over anhydrous sodiumsulphate and concentrated under vacuum. The residue thus obtained waspurified by column chromatography (10-20% EtOAc-Hexane) to afford thetitle compound E28 as a pale yellow solid (0.2 g, 56%). mp 98-100° C.;HPLC: Inertsil ODS 3V (250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄(pH 7.0); solvent B=CH₃CN], Gradient elution program: T/% B=0/60,10/60,25/80, 40/80, 45/60, 50/60; 268 nm, R_(t) 29.79 min, 99.82%purity; MS (CI): m/z 458 (M+H, 100).

Example 118 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-[2-(2-fluorophenoxy)ethoxy]-1,3,5-triazine-4,2-diamine (E29)

[0932]

[0933] A mixture of 2-(2-fluorophenoxy)-1-ethanol (367 mg, 2.356 mmol)and sodium hydroxide (94 mg, 2.356 mmol) in benzene (15 mL) was heatedto reflux for 3 hours with stirring under nitrogen atmosphere and thencooled to 25° C. followed by the addition of compound 1 (0.30 g, 0.785mmol) at same temperature. The mixture was heated to reflux for 12hours, concentrated under vacuum and diluted with water (20 mL). Themixture was then extracted with EtOAc (2×20 mL). The combined organiclayers were washed with water (2×10 mL), dried over anhydrous sodiumsulphate and concentrated under vacuum. The residue thus obtained waspurified by column chromatography (10-20% EtOAc-Hexane) to afford thetitle compound E29 as a semi solid (90 mg, 23%). HPLC: Inertsil ODS 3V(250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄ (pH 7.0); solventB=CH₃CN], Gradient elution program: T/% B=0/60, 10/60,25/80, 40/80,45/60, 50/60; 268 nm, R_(t) 32.31 min, 99.30% purity; MS (CI): m/z 502(M+H, 100).

Example 119 Synthesis ofN²-(3-chloro-4-methoxyphenyl)-N⁴-cycloheptyl-6-(6-methyl-2-pyrylmethoxy]-1,3,5-triazine-4,2-diamine (E30)

[0934]

[0935] A mixture of 6-methyl-2-pyridylmethanol (145 mg, 1.17 mmol) andsodium hydroxide (63 mg, 1.57 mmol) in benzene (15 mL) was heated toreflux for 3 hours with stirring under nitrogen atmosphere and thencooled to 25° C. followed by the addition of compound 133 (0.30 g, 0.785mmol) at same temperature. The mixture was heated to reflux for 12hours, concentrated under vacuum and diluted with water (20 mL). Themixture was then extracted with EtOAc (2×20 mL). The combined organiclayers were washed with water (2×10 mL), dried over anhydrous sodiumsulphate and concentrated under vacuum. The residue thus obtained waspurified by column chromatography (70-80% EtOAc-Hexane) to afford thetitle compound E30 as a hygroscopic solid (257 mg, 70%). HPLC: InertsilODS 3V (250×4.6 mm) 5 microns [solvent A=0.01 M KH₂PO₄ (pH 7.0); solventB=CH₃CN], Gradient elution program: T/% B=0/60, 10/60, 25/80, 40/80,45/60, 50/60; 268 nm, R_(t) 23.93 min, 99.14% purity; MS (CI): m/z 469(M⁺, 100).

Example 120 Synthesis ofN-(3-chloro-4-methoxypheyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine (137)

[0936]N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine (137) is a 1,3,5-triazinederivative which is synthesized from 2,4,6-trichloro-1,3,5-triazine(cyanuric chloride) by serial substitution of amines, for example,3-chloro-4-methoxyaniline, cycloheptylamine, and1-methyl-(4-methylamino)piperidine, respectively.

[0937] This reaction scheme illustrates synthesis of compound 137 byreaction of cyanuric chloride with 3-chloro-4-methoxyaniline usingsodium hydroxide (NaOH) as base at 0-5° C. to yield (3-chloro-4-methoxyphenyl)-(4,6-dichloro-[1,3,5]triazine-2-yl) amine (compound 101), whichon reaction with cycloheptylamine in presence of NaOH under reflux gave6-chloro-N-(3-Chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(compound 133). Compound 133 on reaction with1-methyl-(4-methylamino)piperidine yields, the final compound 137.

Example 121 Alternative Synthesis ofN-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine (137)

[0938]

[0939] A 1 kg campaign was conducted to develop a process which avoidsmultiple column purifications and employs a suitable recrystallisationmethod to produce compound 137. This scheme was stabilized for thecampaign, however the scale up of compound 133 by one pot synthesisresulted in the formation of an impurity, which was carried over to thefinal step and was difficult to remove. The impurity was isolated andcharacterized to be6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-(3-chloro-4-ethoxyphenyl)[1,3,5]triazine-2,4-diamine.To avoid the impurity, the single pot reaction was avoided and compound101 was isolated and purified by recrystallisation in ethylacetate.

[0940] Attempts to minimize the impurity involved scanning several basesand solvents, but the production6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-(3-chloro-4-methoxyphenyl)[1,3,5]triazine-2,4-diamineremained. The preparation of compound 101 was then carried out withoutaddition of base, resulting in the levels of the impurity beingnegligible. Compound 133 was prepared from compound 101 using aqueousNaOH as base.

Example 122 General Large Scale Synthesis ofN-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine (137)

[0941]

[0942] Among the bases and solvents screened, sodium acetate (NaOAc) and1,4-dioxane showed consistently good results for a 1 kg preparation.This scheme involves the preparation of compound 101 by reaction ofcyanuric chloride with 3-chloro-4-methoxyaniline in acetone at 0-5° C.in the absence of base, which is then made to react withcycloheptylamine in presence of NaOH under reflux to give compound 133.This compound 133 was reacted with 1-methyl-(4-methylamino)piperidineusing NaOAc in 1,4-dioxane solvent to yield the crude compound 137 whichis purified by making hydrochloride salt,N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine, HCl salt, compound E31and by recrystallisation in isopropanol. The hydrochloride salt was thenreleased to give the free base, compound 137. Compound obtained in thismanner exhibited the same high quality of appearance, purity, lowresidual solvents, assay and impurity profile, and the like.

Example 123 Large Scale Synthesis of(3-chloro-4-methoxyphenyl)-(4,6-dichloro-[1,3,5]triazine-2-yl)amine(101)

[0943] As provided in the reaction scheme above, a solution of3-chloro-4-methoxyaniline (42.5 g) in acetone was slowly added to thecyanuric chloride solution (50 g dissolved in acetone at −10-0° C. Thereaction mass was maintained at the same temperature for about 1 hourafter addition. The progress of the reaction was monitored by TLC. Afterthe completion of the reaction, the reaction mass was poured overcrushed ice under stirring. The solid compound thrown out was filteredunder vacuum, washed thoroughly with water and then dried. The abovecompound was recrystallised from ethylacetate (Wt. 60 g Y: 73%).

[0944] The following Table illustrates the results obtained usingvarious bases, solvents, and other reaction conditions. Rxn Yield Temp.Cmpdd Disub Trisubstit No. Base Solvent (%) ° C. Purity(%) (%) (%)Remarks 1 NaOH Acetone 92 0-5 90 4-6 1-2 2 NaOH Acetone 75  5-10 86 7-82-3 3 NaOH Acetone 80 0-5 40 35 20 Reverse addition 4 NaOH Acetone 85 3065 15-20 5-8 5 NaOH Acetone 89 Reflux 20 60 10-15 6 K₂CO₃ Acetone 90 0-591 7-8 1-2 7 K₂CO₃ Toluene 90 0-5 91 7-8 1-2 8 NaOH Toluene 80 0-5 856-7 1-2 9 NaOH Dioxane 85 0-5 92 5-6 — 10 K₂CO₃ Dioxane 90 0-5 91 5-6 —11 K₂CO₃ Dioxane 85 0-5 55 20-25 4-5 Reverse addition 12 — Acetone 950-5 95 1-2 — 13 — Acetone 98 <0 99 — — 14 Basic Acetone 85 0-5 95 — —resin

Example 124 Large Scale Synthesis of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(133)

[0945] As provided in the reaction scheme above, a solution ofcycloheptylamine 20.4 g) in acetone was slowly added to the 101 solution(50 g dissolved in acetone) at 20-30° C. To this stirring solution water(100 ml) was added followed by Aq. sodium hydroxide solution (6.5 gdissolved in water). Then the temperature of the reaction mass raised toreflux and maintained for about 2-3 h. TLC was monitored. Aftercompletion of the reaction, temperature of the RM (reaction mixture) wasbrought to 25-30° C. and the reaction mass was poured over chilled waterunder stirring. The solid compound thrown out was filtered and dried(Wt. 60 g Y: 95.8%).

[0946] The following Table illustrates the results obtained usingvarious bases, vents, and other reaction conditions. Rxn Yield Cmpd No.Base Solvent (%) Purity(%) Remarks 1 NaOH Acetone 92 95 2 NaOH Toluene80 90 3 NaOH Dioxane 80 82 4 K₂CO₃ Acetone 95 99 5 K₂CO₃ Dioxane 89 85 6Et₃N Acetone 85 75 7 Et₃N Dioxane 75 60 8 Et₃N Toluene 70 65

Example 125 One-Pot Synthesis of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(133)

[0947]

[0948] A convenient, one-pot synthesis of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diaminewas carried out as follows. To cyanuric chloride (50 g) dissolved inacetone was added a solution of 3-chloro-4-methoxyaniline (42.5g) inacetone at 0-5° C. followed by the addition of potassium carbonate (112g). The reaction mass was maintained at the same temperature for about1hour. The progress of the reaction was monitored by TLC. When thereaction was complete, cycloheptylamine (29 g dissolved in acetone) wasadded at 0-10° C. The reaction mass was maintained under reflux forabout 2-3hours. Completion of the reaction was checked by TLC. Thereaction mass was cooled to 20-30° C. and poured over crushed ice understirring. The solid compound thrown out was filtered and dried. Thecrude compound was then recrystallised from ethylacetate (Wt.50 gY:48%).

Example 126 Large-Scale Synthesis ofN-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methylpiperidin-4-yl)[1,3,5]triazine-2,4,6-triamine (137)

[0949] 1-Methyl-(4-methylamino)piperidine (17 g in 1,4-dioxane) wasadded to the solution of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(50 g dissolved in 1,4-dioxane) at RT followed by addition of sodiumacetate (21.5 g). The RM (reaction mixture) was maintained under refluxfor about 2-3 hours. The progress of the reaction was monitored by TLC.After the completion of the reaction, the reaction mass was cooled to20-30° C. and filtered through hi-flow bed. The filtrate was evaporatedto minimum volume and was diluted with toluene. This was acidified withdilute HCl under stirring conditions. The solid obtained was filteredand dried. The above crude compound was recrystallised from isopropanol(Wt. 20 g Y:30%).

[0950] The HCl salt (20 g) was dissolved in methanol, diluted withethylacetate and made basic with aqueous potassium carbonate solution.The organic layer was separated and the aqueous layer was againextracted into ethylacetate. The organic layers were combined, washedwith water and evaporated to yield the compound (Wt. 20 g Y: 30%).

Example 127 General Synthesis of Akoxide Derivatives of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine

[0951]

[0952] The sequential substitution at the triazine core permitted theconvenient formation of alkoxide derivatives of triazines. The procedureinvolves the preparation of compound 101 by reaction of cyanuricchloride with 3-chloro-4-methoxyaniline in acetone at 0-5° C. in theabsence of base, which is then made to react with cycloheptylamine inpresence of sodium hydroxide under reflux to give compound 133. Thiscompound 133 was refluxed in alcohols like methanol, ethanol,isopropanol, etc., with base to yield alkoxide compounds whosestructures are confirmed by the spectral data.

Example 128 Synthesis of 6-methoxy-N-(3-Chloro-4-methoxyphenyl)-N′-cyclo heptyl-[1,3,5]triazine-2,4diamine (E32)

[0953] To6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(5 g) dissolved in methanol was added K₂CO₃ (3.2 g) and refluxed forabout 6-8 h. TLC was monitored. After the completion of the reaction,the reaction mass was filtered through vacuum and the filtrate wasevaporated to yield the title compound. (Wt. 4.7 g Y: 95%)

Example 129 Synthesis of 6-ethoxy-N-(3-Chloro-4-methoxy phenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4diamine (E33)

[0954] A portion of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(5 g) was refluxed in ethanol in presence of K₂CO₃ (3.2 g) for about8-10 h. TLC was monitored. After the completion of the reaction, thereaction mass was filtered through vacuum and the filtrate wasevaporated to yield the title compound. (Wt. 4.6 g Y: 90%).

Example 130 Synthesis of 6-isopropoxy-N-(3-Chloro-4-methoxyphenyl)-N′-cyclo heptyl-[1,3,5]triazine-2,4diamine (E34)

[0955] To a portion of6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(5 g) in isopropanol was added K₂CO₃ (3.2 g) and refluxed for about10-12 h. TLC was monitored. After the completion of the reaction, thereaction mass was filtered through vacuum and the filtrate wasevaporated to yield the title compound. (Wt. 3.7 g Y: 70%).

Example 131 Alternative Synthesis of 6-alkoxy-N-(3-Chloro-4-methoxyphenyl)-N′cyclo heptyl-[1,3,5]triazine-2,4diamine

[0956]

[0957] To6-chloro-N-(3-chloro-4-methoxyphenyl)-N′-cycloheptyl-[1,3,5]triazine-2,4-diamine(5 g) dissolved in solvent (Methanol, Ethanol, Isopropanol) was addedK₂CO₃ (3.2 g) and refluxed for about 8-10 h. TLC was monitored. Aftercompletion of the reaction, the reaction mass was filtered throughvacuum and the filtrate was evaporated to yield the title compounds,E32, E33, and E34, respectively. Rxn No. Solvent Yield (%) Purity (%) 1Methanol 90 95 2 Ethanol 85 96 3 Isopropanol 50 80

Example 132 Biological Activity Data

[0958] Additional bioactivity data for compounds of the presentinvention are presented in Table 8, where compounds that have at leastthe activity of effecting cellular proliferation or at least theactivity of modulating inflammation activity, as measured by the assaystaught herein, are provided. For compounds that show at least theactivity of modulating inflammation activity, the activity is typicallymeasured and reported relative to IL6 production compared to cells thatdid not receive compound (or per cent of control IL6 production). Theinclusion of compounds in the categories of the Tables disclosed herein,including this Table, is not to be seen as limiting, in that compoundsincluded in such Tables have at least the activity shown for inclusionin the Table and may have more or other activities. Nor are the Tablesto be seen as limiting in that these are the only compounds disclosedherein that have that activity, representative compounds are shown inthe Tables that have at least that particular activity for inclusion inthe Table. One or more compounds disclosed herein have at least anactivity that has utility in treatment of disease states. Note that anyhydrogen atoms that are required for any atom to attain its usualvalence in a structure presented in Table 8, whether a carbon atom or aheteroatom, should be inferred if it is not specifically indicated.

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[1017] 59. Janietz et al., SYNTHESIS, 33-34 (1993) TABLE 1DRepresentative Compounds of the Present Invention Cmpd Structure Name D1

6-(3-Chloro-4-methoxy-phenoxy)-N-cyclo-heptyl-N′-methyl-N′-(1-methyl-piper- idin-4-yl)-]1,3,5]triazine-2,4-di-amine D2

N-Cycloheptyl-6-(3-fluoro-4-methoxy-phenoxy)-N′-methyl-N′-(1-methyl-piper-idin-4-yl)-[1,3,5]triazine-2,4-di- amine D3

N-(3-Chloro-4-methoxy-phenyl)-6-cyclo- heptyloxy-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4- diamine D4

6-Cycloheptyloxy-N-(3-fluoro-4-meth- oxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4- diamine D5

N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-(1-methyl-piperidin-4-yl- oxy)-[1,3,5]triazine-2,4-diamine D6

N-(3-Chloro-4-methoxy-phenyl)-N′-cyclo-heptyl-6-(1-methyl-piperidin-4-yl- oxy)-[1,3,5]triazine-2,4-diamine D7

N-Cycloheptyl-N′-(3-fluoro-4-methoxy- phenyl)-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine D8

N-(3-Chloro-4-methoxy-phenyl)-N′-cyclo- heptyl-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine D9

(3-Chloro-4-methoxy-phenyl)-[4-cyclo- phenyloxy-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-amine D10

[4-Cycloheptyloxy-6-(1-methyl-piper- idin-4-yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine D11

N-Cycloheptyl-N′-(1-ethyl-pyrrol-idin-2-ylmethyl)-6-(3-fluoro-4-methoxy- phenoxy)-[1,3,5]triazine-2,4-di-amine D12

6-(3-Chloro-4-methoxy-phenoxy)-N- cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4-di- amine D13

6-Cycloheptyloxy-N-(1-ethyl-pyrrol-idin-2-ylmethyl)-NN′-(3-fluoro-4-meth-oxy-phenyl)-[1,3,5]triazine-2,4-di- amine D14

N-(3-Chloro-4-methoxy-phenyl)-6-cyclo- heptyloxy-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4-di- amine D15

N-Cycloheptyl-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-N′-(3-fluoro-4-methoxy- phenyl)-[1,3,5]triazine-2,4-di-amine D16

N-(3-Chloro-4-methoxy-phenyl)-N′-cyclo- heptyl-6-(2-ethyl-pyrrolidin-2-ylmethoxy)-[1,3,5]triazine-2,4-diamine D17

[4-Cycloheptyloxy-6-(3-fluoro-4-meth- oxy-phenoxy)-[1,3,5]triazin-2-yl]-methyl-(1-methyl-piperidin-4-yl)- amine D18

[4-Cycloheptyloxy-6-(piperidin-4- yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine D19

Cycloheptyl-[4-(3-fluoro-4-methoxy- phenoxy)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl)-amine D20

2-Cycloheptyloxy-4-(3-fluoro-4-meth- oxy-phenoxy)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine D21

2-[3-Chloro-4-methoxy-phenoxy)-4- cycloheptyloxy-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine D22

[4-Cycloheptyloxy-6-(3-fluoro-4- methoxy-phenoxy)-[1,3,5]triazin-2-yl]-(1-ethyl-pyrrolidin-2-ylmethyl)- amine D23

[4-Cycloheptyloxy-6-(1-ethyl-pyrrol- idin-2-ylmethoxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)- amine D24

Cycloheptyl[4-(1-ethyl-pyrrolidin- 2-ylmethoxy)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]-amine D25

2-Cycloheptyloxy-4-(1-ethyl-pyrrol-idin-2-ylmethoxy)-6-(3-fluoro-4-meth- oxy-phenoxy)-[1,3,5]triazine D26

N-(4-Amino-3-chloro-phenyl)-N′-cyclo- heptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6- triamine D27

1-{4-[4-Cycloheptylamino-6-(3- fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piper- idin-2-yl}-ethanone D28

4-[4-Cycloheptylamino-6-(3-fluoro-4- methoxy-phenylamino)-[1,3,5]tri-azin-2-ylamino]-piperidine-1-carboxy- lic acid D29

4-[4-Cycloheptylamino-6-(3-fluoro-4- methoxy-phenylamino)-[1,3,5]tri-azin-2-ylamino]-piperidine-1-carboxy- lic acid amide D30

4-[4-Cycloheptylamino-6-(3-fluoro-4- methoxy-phenylamino)-[1,3,5]tri-azin-2-ylamino]-piperidin-1-ol D31

2-{[4-Cycloheptylamino-6-(3-fluoro- 4-methoxy-phenylamino)-[1,3,5]tri-azin-2-ylamino]-methyl}-piperidine-1- carboxylic acid D32

2-{[4-Cycloheptylamino-6-(3-fluoro- 4-methoxy-phenylamino)-[1,3,5]tri-azin-2-ylamino]-methyl}-pyrrolidine-1- carboxylic acid amide D33

Acetic acid 2-chloro-4-{4-cycloheptyl- amino-6-[methyl-(1-methyl-piper-idin-4-yl)-amino]-[1,3,5]triazin-2-yl- amino]phenyl ester D34

N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-(tetrahydro-pyran-4-yl)- [1,3,5]triazine-2,4,6-triamine D35

N-Cycloheptyl-N′-(3-fluoro-4-meth- oxy-phenyl)-N″-methyl-N″-(tetrahydro-pyran-4-yl)-[1,3,5]triazine-2,4,6- triamine D36

4-[4-Cycloheptylamino-6-(piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]- 2-fluoro-phenol D37

2-Chloro-4-[4-cycloheptylamino-6- (piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]-phenol D38

N-(4-Aminomethyl-cyclohexyl)-N′-cyclo- heptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-tri- amine D39

N-(4-Aminomethyl-cyclohexyl)-N′-(3- chloro-4-methoxy-phenyl)-N″cyclo-heptyl-[1,3,5]triazine-2,4,6-tri- amine D40

4-[4-(4-Aminomethyl-cyclohexylamino)- 6-cycloheptylamino-[1,3,5]triazin-2-ylamino]-2-fluoro-phenol D41

N-CycloheptylN′-(3-fluoro-4-meth- oxy-phenyl)-N″-piperidin-4-ylmethyl-[1,3,5]triazine-2,4,6-triamine D42

N-(3-Chloro-4-methoxy-phenyl)-N′- cycloheptyl-N″-piperidin-4-ylmethyl-[1,3,5]triazine-2,4,6-triamine D43

4-{4-Cycloheptylamino-6-[(piperidin- 4-ylmethyl)-amino]-[1,3,5]triazin-2-ylamino}-2-fluoro-phenol D44

N-Cycloheptyl-N′-methyl-N′-(1-methyl- piperidin-4-yl)-[1,3,5]triazine-2,4,6,-triamine D45

N-Cycloheptyl-N′-piperidin-4-yl[1, 3,5]triazine-2,4,6-triamine D46

N-(3-Fluoro-4-methoxy-phenyl)-N′- methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine D47

4-(3-Fluoro-4-methoxy-phenylamino)- 6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol D48

4-(3-Chloro-4-methoxy-phenylamino)- 6-[methyl-(1-methyl-pipeidin-4-yl)-amino]-[1,3,5]triazin-2-ol D49

4-(3-Chloro-4-methoxy-phenylamino)- 6-cycloheptylamino-[1,3,5]triazin-2-ol D50

4-Cycloheptylamino-6-(3-fluoro-4- methoxy-phenylamino)-[1,3,5]triazin-2-ol D51

4-Cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol D52

N-(4-Amino-cyclohexylmethyl)-N′-(3- chloro-4-methoxy-phenyl)-N″-cyclo-heptyl-[1,3,5]triazine-2,4,6-triamine D53

N-(4-Amino-cyclohexylmethyl)-N′- cycloheptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine D54

4-Cycloheptylamino-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol D55

4-[(1-Ethyl-pyrrolidin-2-ylmethyl)- amino]-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol D56

4-(3-Chloro-4-methoxy-phenylamino)- 6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol D57

4-(Cyclohexylmethyl-amino)-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol D58

4-(Cyclohexylmethyl-amino)-6-[(1- ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol

[1018] TABLE 2 Representative compounds of the present inventionprepared by parallel synthesis reactions, including amine monomersproducts, and characterization data Cmpd No. R1 (Monomer 1) R2 (Monomer2) R3 (Monomer 3) 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

LC-MS (Calc'd MW) Synthetic Observed MW Cmpd No. Product (StructureProduct Name Procedure (M + H) 1

N2-(4-bromo-1-naphthyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (538) 540.4 2

N2-(4-chloro-1-naphthyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (493) 494.3 3

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-quinolinyl)-1,3,5-triazine-2,4,6- triamine Parallel synthesis method A(460) 461.5 4

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(6-quinolinyl)-1,3,5-triazine-2,4,6- triamine Parallel synthesis method A(460) 461.4 5

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(6-quinolinyl)-1,3,5-triazine-2,4,6- triamine Parallel synthesis method A(460) 461.4 6

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[1-(2-naphthyl)ethyl]-1,3,5-triazine- 2,4,6-triamine Parallel synthesis methodA (487) 488.5 7

N2-cycloheptyl-N4-(3,4- dichlorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (478) 478.3 8

N2-cycloheptyl-N4-(3,4- difluorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A 445 (446.3) 9

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(trifluoromethoxy)phenyl]-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method A (493) 494.3 10

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-fluorophenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(493) 428.4 11

4-[(4-(cycloheptylamino)-6- {[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazine-2-yl)amino]benzonitrileParallel synthesis method A (434) 435.4 12

N2-(4-chlorophenyl-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (444) 444.3 13

N2-(4-bromophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (488) 488.4 14

4-[(4-(cycloheptylamino)-6-{[(1- ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazin-2-yl)-amino]benzoate Parallelsynthesis method A (481) 482.5 15

N2-(1,1′-biphenyl-4-yl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (485) 486.5 16

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluorophenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(427) 428.3 17

N2-(3-chlorophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (444) 444.3 18

N2-(3-bromophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (488) 488.4 19

3-[(4-cycloheptylamino)-6-{[(1- ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazine-2-yl)-amino]benzoate Parallelsynthesis method A (481) 482.4 20

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-fluorophenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(427) 428.4 21

N2-(2-chlorophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (444) 444.3 22

N2-(2-bromophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (488) 490.3 23

N2-(1,3-benzodioxol-5-yl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Paralell synthesismethod A (453) 454.4 24

N2-cycloheptyl-N4-(2,3-dihydro- 1,4-benzodioxin-6-yl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (467) 454.4 25

N2-cycloheptyl-N4-[4- (dimethylamino)phenyl]-N6-[(1-ethyl-2-pyrrolidinyl)methyl]- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (452) 453.5 26

N2-[3-chloro-4- (diethylamino)phenyl-N4- cycloheptyl-N6-[1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (515) 515.5 27

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(4-morpholino)phenyl[-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (494) 495.4 28

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(4-methyl-1-piperazinyl)phenyl]- 1,3,5-triazine-2,4,60triamine Parallelsynthesis method A (507) 508.5 29

N-{4-[(4-(cycloheptylamino)-6- {[(1-ethyl-2-pyrrolidinyl)methyl]amino)-1,3,5- triazin-2-yl)- amino]phenyl}acetamideParallel synthesis method A (466) 467.3 30

N-(3-[(4-cycloheptylamino-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazin-2-yl)- amino]phenyl}acetamideParallel synthesis method A (466) 467.4 31

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-methoxyphenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis methodA (439) 440.4 32

N2-cycloheptyl-N4-(4- ethoxyphenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (453) 454.2 33

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(methylthio)phenyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (455) 456.4 34

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-pyridinyl)-1,3,5-triazine-2,4,6- triamine Parallel synthesis method A(410) 411.3 35

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-methylphenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(423) 36

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-phenoxyphenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis methodA (501) 502.3 37

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-methylphenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(423) 424.4 38

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-methylphenyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis method A(423) 424.3 39

2-[(4-cycloheptylamino)-6-{[(1- ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazin-2-yl)-amino]-4-methyl-3-thiophenecarboxamide Parallel synthesis method A (472) 473.3 40

N2-(4-chlorophenyl)-N4- cyclohpetyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N2-methyl- 1,3,5-triazine-2,4,6-triamine Paralellsynthesis method A (458) 458.5 41

3-[(4-(cycloheptylamino)-6-{[(1- ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazine-2-yl)-(phenyl)amino]propanenitrile Parallel synthesis method A (462) 463.4 42

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-methoxyphenyl)-N6-methyl- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (543) 454.4 43

N2-cycloheptyl-N4-(2,4- difluorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-methyl- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (459) 460.3 44

[(4-(cycloheptylamino)-6-{[(1- ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazine-2-yl)(phenyl)amino]acetonitrile Parallel synthesis method A (448) 449.3 45

N2-(3-chlorophenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N2-methyl- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (458) 458.5 46

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-methyl-N6-[2-(trifluorometnhyl)phenyl]- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (491) 492.3 47

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-methyl-N6-[2-(trifluoromethoxy)phenyl]- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method A (507) 508.3 48

N2-(3-chloro-4-methoxyphenyl)- N4-cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod A (474) 474.3 49

N-benzoyl-4-[(3- (cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}- 1,3,5-triazine-2-yl)-amino]benzenesulfonamide Parallel synthesis method A (592) 593.4 50

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-naphthyl)-1,3,5-triazine-2,4,6- triamine Parallel synthesis method A(459) 460.4 51

N²ethyl-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (389) 390.2 52

N²-(tert-butyl)-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (417) 418.4 53

N²-benzyl-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (451) 452.3 54

N²-cyclooctyl-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (471) 472.4 55

N²-cyclohexyl-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-0 triazine-2,4,6-triamine Parallelsynthesis method B (443) 444.3 56

N²-cycloheptyl-N⁴-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (429) 430.3 57

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-(1- pyrrolidinyl)-1,3,5-triazine-2,4- diamineParallel synthesis method B (415) 416.3 58

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3- fluoro-4-methoxyphenyl)-6-hexahydro-1H-azepin-1-yl- 1,3,5-triazine-2,4-diamine Parallel synthesismethod B (443) 444.4 59

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3- fluoro-4-methoxyphenyl)-6-oxtahydro-1(2H)-quinolinyl- 1,3,5-triazine-2,4-diamine Parallelsynthesis method B (483) 484.5 60

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(4- methylcyclohexyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method B 457 (458.4) 61

N²-(1-ethyl-pyrrolidin-2- ylmethyl]-N⁴-(3-fluoro-4-methoxyphenyl)-6-((S)-2- methoxymethyl-pyrrolidin-1-yl)-1,3,5-triazine-2,4-diamine Parallel synthesis method B (459) 460.4 62

N²-[(1-ethyl-2- pyrrolidin)methyl]-N⁴-(3- fluoro-4-methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod B (444) 445.3 63

6-(4-acetyl-1-piperazinyl)-N²- [(1-ethyl-2-pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)- 1,3,5-triazine-2,4-diamine Parallelsynthesis method B 472 (473.3) 64

Ethyl-4-{4-{[(1-ethyl-2- pyrrolidinyl)methyl]amino}-N⁴- [(3-fluoro-4-methoxyphenyl)-amino]-1,3,5- triazin-2-yl)-1- piperazinecarboxylateParallel synthesis method B (502) 503.2 65

N²-(cyclohexylmethyl)-N⁴-[(1- ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method B (457) 458.4 66

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶(2- furylmethyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method B (441) 442.2 67

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3- fluoro-4-methoxyphentl)-N⁶-(2,2,2-trifluoroethyl)-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod B (443) 444.1 68

N²-[2-(dimethylamino)-ethyl]-N⁴- [(1-ethyl-2-pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)- 1,3,5-triazine-2,4,6-triamine Parallelsynthesis method B (432) 433.5 69

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-(4- [2-oxo-2-(1-pyrrolidinyl)ethyl]-1-piperazinyl)-1,3,5-triazine-2,4- diamine Parallel synthesis method B(541) 542.4 70

N²,N⁴-bis[(1-ethyl-2- pyrrolidinyl)methyl]-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method B (472) 473.3 71

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[2- (1-piperidinyl)ethyl]-1,3,5-triazine-2,4,6-triamine Parallel synthesis method B (472) 473.3 72

N²-[4-(1,3-benzodioxol-5- ylmethyl)-1-piperazinyl]-N⁴-[(1-ethyl-2-pyrrolidinyl)methyl]-N⁴(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine Parallel synthesis method B (564) 565.4 73

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[4- (2-pyridyl)-1-piperazinyl]-1,3,5-triazine-2,4-diamine Parallel synthesis method B (507) 508.3 74

1-[3-({4-{[(1-ethyl-2- pyrrolidinyl)methyl]amino}-6-[(3-fluoro-4-methoxyphenyl)amino}- 1,3,5-triazin-2-yl)amino)propyl]-2-pyrrolidone Parallel synthesis method B (486) 487.5 75

N²-[(1-ethyl-2- pyrrolidinyl)methyl]-N⁴-(3-fluoro-4-methoxyphenyl)-N⁶-[3- (1H-imidazol-1-yl)propyl]-1,3,5-triazine-2,4,6-triamine Parallel synthesis method B (469) 470.3 76

N2-cycloheptyl-N4-ethyl-N6-(3- fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (374) 375.3 77

N2-(tert-butyl)-N4-cycloheptyl- N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (402) 403.4 78

N2-benzyl-N4-cycloheptyl-N6- (3-fluoro-4-methoxyphenyl)1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (436) 436.9 79

N2-cycloheptyl-N4-cyclooctyl- N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (456) 457.3 80

N2-cycloheptyl-N4-cyclohexyl- N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (428) 429.3 81

N2-cycloheptyl-N4-cyclopentyl- N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6-triamine Parallel synthesis method C (414) 415.2 82

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-(1-pyrrolidinyl)-1,3,5-triazine-2,4- diamine Parallel synthesis method C(400) 400.9 83

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-hexahydro-1H-azepin-1-yl-1,3,5-triazine- 2,4-diamine Parallel synthesis method C(428) 429 84

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-octahydro-1(2H)-quinolinyl-1,3,5-triazine- 2,4-diamine Parallel synthesis method C(468) 469.3 85

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-(4-methylcyclohexyl)-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod C (442) 443.3 86

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-[(2S)-2-(methoxymethyl)-1-pyrrolidinyl]- 1,3,5-triazine-2,4-diamine Parallelsynthesis method C (444) 444.9 87

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5-triazine-2,4- diamine Parallel synthesis method C(429) 430.3 88

6-(4-acetyl-1-piperazinyl)-N2- cycloheptyl-N4_93-fluoro-4-methoxyphenyl)-1,3,5-triazine- 2,4-diamine Parallel synthesis method C(457) Wrong MW observed 89

ethyl-4-(4-(cycloheptlyamino)-6- [(3-fluoro-4-methoxyphenyl)amino]-1,3,5- triazin-2-yl]-1- piperazinecarboxylateParallel synthesis method C (487) 488.3 90

N2-cycloheptyl-N4- (cyclohexylmethyl)-N6-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod C (442) 91

N2-cycloheptyl-N4-(3-fluoro-4- methoxy)phenyl)-N6-(2-furanylmethyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesis methodC (426) 426.9 92

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-(2,2,2-trifluoromethyl)-1,3,5-triazine- 2,4,6-triamine Parallel synthesismethod C (428) 352.1 93

N2-cycloheptyl-N4-[2- (dimethylamino)ethyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method C (417) 418.3 94

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-{4-[2-oxo-(1-pyrridinyl)ethyl -1-piperazinyl)- 1,3,5-triazine-2,4-diamine Parallelsynthesis method C (526) 527.3 95

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphentl)-1,3,5- triazine-2,4,6-triamine Parallelsynthesis method C (457) 456.4 96

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-[2-(1-piperidinyl)ethyl]-1,3,5-triazine- 2,4,6-triamine Parallel synthesismethod C (457) 458.4 97

6-[4-(1,3-benzodioxol-5- ylmethyl)-1-piperazinyl]-N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-1,3,5-triazine- 2,4-diamineParallel synthesis method C (549) 493.2 98

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-[4-(2-pyridinyl)-1-piperazinyl]-1,3,5- triazine-2,4-triamine Parallelsynthesis method C (492) incorrect MW observed 99

1-[3-((4-(cycloheptylamino)-6- [(3-fluoro-4-methoxyphenytl)amino]-1,3,5- triazine-2-yl)amino)propyl]-2-pyrrolidinone Parallel synthesis method C (471) 472.3 100

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-[3-(1H-imidazol-1-yl)propyl]-1,3,5- triazine-2,4,6-triamine Parallel synthesismethod C (454) 455.3 Yield Based on Purification Cmpd No. LC PurityPurity method 1 62 23 ISCO 2 54 18 ISCO 3 95 25 ISCO 4 97 26 ISCO 5 9861 ISCO 6 94 39 ISCO 7 94 53 ISCO 8 98 55 ISCO 9 96 40 ISCO 10 90 37ISCO 11 76 64 none 12 76 69 none 13 77 70 none 14 96 46 ISCO 15 75 69none 16 78 75 none 17 77 74 none 18 82 75 none 19 98 45 ISCO 20 60 23ISCO 21 53 26 ISCO 22 51 17 ISCO 23 16 4 ISCO 24 91 30 ISCO 25 67 17ISCO 26 91 45 ISCO 27 92 41 ISCO 28 4 0.004 none 29 91 26 ISCO 30 100 38ISCO 31 96 42 ISCO 32 97 47 ISCO 33 93 49 ISCO 34 27 7 ISCO 35 72 76none 36 100 46 ISCO 37 86 46 ISCO 38 98 44 ISCO 39 63 16 ISCO 40 98 42ISCO 41 97 45 ISCO 42 89 36 ISCO 43 73 74 none 44 85 76 none 45 98 37ISCO 46 82 21 ISCO 47 100 41 ISCO 48 93 51 ISCO 49 14 2 none 50 83 36ISCO 51 48 36 none 52 36 28 none 53 87 75 none 54 83 66 none 55 75 80none 56 79 75 none 57 84 45 ISCO 58 90 51 ISCO 59 86 47 ISCO 60 88 50ISCO 61 61 36 ISCO 62 30 10 none 63 83 43 ISCO 64 75 74 none 65 76 77none 66 77 70 none 67 77 69 none 68 6 2 none 69 78 39 ISCO 70 63 20 ISCO71 67 28 ISCO 72 87 83 none 73 86 41 ISCO 74 85 31 ISCO 75 90 38 ISCO 7685 18 preparative HPLC 77 81 3 preparative HPLC 78 98 15 preparativeHPLC 79 96 5 preparative HPLC 80 100 10 preparative HPLC 81 89 26preparative HPLC 82 98 22 preparative HPLC 83 96 5 preparative HPLC 8467 21 preparative HPLC 85 64 4 preparative HPLC 86 100 17 preparativeHPLC 87 94 13 preparative HPLC 88 100 compound not preparative observedafter HPLC prep HPLC 89 97 4 preparative HPLC 90 100 16 preparative HPLC91 59 31 preparative HPLC 92 93 1 preparative HPLC 93 76 11 preparativeHPLC 94 27 15 preparative HPLC 95 80 24 preparative HPLC 96 86 21preparative HPLC 97 94 21 preparative HPLC 98 70 compound notpreparative observed after HPLC prep HPLC 99 96 29 preparative HPLC 10094 24 preparative HPLC

[1019] TABLE 3 Triazine compounds active in anti-proliferation assay(perlecan), generally useful in treating cardiovascular disordersassociated with Activity Patent # STRUCTURE CAS NAME (note 1) Avg IC-50100

HCl N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triaminehydrogen chloride ++ 4-5 95

N2-cycloheptyl-N4-(1-ethyl-pyrrolidin-2-ylmethyl)N6-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine- 2,4,6-triamine + 7.3 26

N2-(3-chloro-4-diethylamino-phenyl)-N4-cycloheptyl-N6-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine + 7.73 139

N2-cycloheptyl-N4-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxy-phenyl)-1,3,5- triazine-2,4,6-triamine++ 4-5 140

N2-cycloheptyl-N4-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N6-(3-fluoro-4-methoxy-phenyl)-1,3,5- triazine-2,4,6-triamine++ 4-5 163

HCl N2-(3-chloro-4-diethylamino-phenyl)-N4-cycloheptyl-N6-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine hydrogen chloride + 8-10 123

N2-(3-bromo-methoxy-phenyl)-N4-cycloheptyl-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triamine +170

HCl N2-(3-bromo-methoxy-phenyl)-N4-cycloheptyl-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triaminehydrogen chloride + 8-9 137

N2-(3-chloro-methoxy-phenyl)-N4-cycloheptyl-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triamine ++4-5 103

N2-(3-chloro-4-methoxy-phenyl)-N4-cyclohexylmethyl-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine + −7 uM 105

N2-(3-chloro-4-methoxy-phenyl)-N4-methyl-N4-(1-methyl-piperidin-4-yl)-N6-(1-propyl-butyl)-1,3,5-triazine-2,4,6-triamine + −7 uM 148

N2-(1-aza-bicyclo[2.2.2]oct-3-yl)-N4-(3-chloro-4-methoxy-phenyl)-N6-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine ++ less than 5 uM 146

N²-(3-chloro-4-methoxy-phenyl)-N⁴-cycloheptyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6- triamine +++ −2.2 138

2-chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl-amino}-1,3,5-triazin-2- ylamino}-phenol ++ 4.03 uM135

N2-(3-chloro-4-methoxy-phenyl)-N4-cycloheptyl-N6-piperidin-4-yl-1,3,5-triazine-2,4,6-triamine 169

N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triaminemaleate + 7.2 uM 167

N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5- triazine-2,4,6-triaminemaleate + 6.8 uM 159

N²-Cycloheptyl-N⁴-(3-fluoro-4-methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5-triazine-2,4,6- triamine ++ 5.7-6.9 uM125

6-chloro-N-cyclohexylmethyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine + 10 uM 127

6-Chloro-N-cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine + 10 uM

[1020] TABLE 4A Triazine compounds that are toxic or cause cell deathfor use in oncology applications Patent Name Number STRUCTURE Generatedby Autonom ™ Toxicity 93

N2-cycloheptyl-N4-[2- (dimethylamino)ethyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine toxic 3

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-quinolinyl)-1,3,5-triazine-2,4,6-triamine toxic 5

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(8-quinolinyl)-1,3,5-triazine-2,4,6-triamine toxic 6

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[1-(2-naphthyl)ethyl]-1,3,5-triazine-2,4,6- triamine toxic 7

N2-cycloheptyl-N4-(3,4- dichlorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine- 2,4,6-triamine toxic 8

N2-cycloheptyl-N4-(3,4- difluorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine- 2,4,6-triamine toxic 9

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(triflouromethoxy)phenyl]-1,3,5- triazine-2,4,6-triamine toxic 12

N2-(4-chlorophenyl)-N4-cycloheptyl- N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine toxic 13

N2-(4-bromophenyl)-N4-cycloheptyl- N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine toxic 14

4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5- triazin-2-yl)-amino]benzoate toxic16

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluorophenyl)-1,3,5-triazine-2,4,6- triamine toxic 21

N2-(2-chlorophenyl)-N4-cycloheptyl- N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine toxic 22

N2-(2-bromophenyl)-N4-cycloheptyl- N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6-triamine toxic 36

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-phenoxyphenyl)-1,3,5-triazine-2,4,6- triamine toxic 47

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-methyl-N6-[2-(trifluoromethoxy)phenyl]-1,3,5- triazine-2,4,6-triamine toxic 54

N2-cyclooctyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-fluoro-3-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine toxic 59

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4- (4-fluoro-3-methoxyphenyl)-6-octahydro-1(2H)-quinolinyl-1,3,5- triazine-2,4-diamine toxic 121

N,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(1-methyl-piperidin-4- yl)-[1,3,5]]triazine-2,4,6-triamineToxic by toxicity assay at 0.75 uM. Starts blunting by 1 uM and roundingby 5 uM 111

N-Butyl-N′-(3-chloro-4-methoxy- phenyl)-N″-methyl-N″-(1-methyl-piperin-4-yl)-N-propyl-[1,3,5]triazine- 2,4,6-triamine Toxic by eye at50 and 10 uM no blunt/round just death 112

N2-Butyl-N4-(3-chloro-4-methoxy- phenyl)-N6-methyl-N6-piperidin-4-yl-N2-propyl-1,3,5-triazine-2,4,6-triamine Toxic by eye at 50 and 10 uM noblunt/round just death 136

N-(3-Chloro-4-methoxy-phenyl)-N′- cycloheptyl-N″-(1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4,6-triamine Nothing but cell debris at 3 uMvery toxic 119

N2-(3-chloro-4-methoxyphenyl)-6- cyclohexylmethoxy-N4-methyl-N4-(1-methyl-piperidin-4-yl)-1,3,5-triazine- 2,4-diamine toxic

[1021] TABLE 4B Triazine complunds toxic to endothelial cells when AGEstimulated % IL6 production compare to TNF % IL6 production compare toAGE Toxic Toxic stimulated of IL6 (100%). stimulation of IL6 (100%).Compound Name (AGE- (TNF- Compds tested @ 10 uM Compds tested @ 10 uMNumber STRUCTURE Generated by Autonom ™ treated) treated) HAEC, IL6HAEC, IL6 121

N,N′-Bis-(3-chloro-4-methoxy-phenyl)-N″-methyl-N″-(4-methyl-cyclohexyl)- (1,3,5]triazine-2,4,6-triamine toxicn.d. total cell death @ 10 uM (50 @ 5 uM, but some toxicity (˜20%)] n.d.168

N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine citrate salt toxic n.d. total celldeath @ 10 uM [40 @ 5 uM] n.d. 159

N2-Cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-piperidin-4-yl-1,3,5- triazine-2,4,6-triamine toxicn.d. total cell death @ 5 and 10 uM n.d. 167

N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine maleate salt toxic n.d. total celldeath @ 10 uM [25 @ 5 uM] n.d. 169

N2-cycloheptyl-N4-(3-fluoro-4-methoxy-phenyl)-N6-methyl-N6-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine succinate salt toxic n.d. total celldeath @ 10 uM [45 @ 5 uM] n.d. 148

N-(1-Aza-bicyclo[2.2.2]oct-3-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-)1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4,6- triamine toxic n.d. totalcell death @ 10 uM [60 @ 5 uM] n.d. 134

N-(1-benzyl-piperidin-4-yl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]- 2,4,6-triamine toxic n.d. totalcell death @ 10 uM [55 @ 5 uM] n.d. 135

N2-(3-chloro-4-methoxy-phenyl)-N4-cycloheptyl-N6-piperidin-4-yl-1,3,5-triazine- 2,4,6-triamine toxic n.d.total cell death @ 10 uM [60 @ uM] n.d. 138

2-chloro-4-{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl-amino]1,3,5-triazin-2- ylamino}-phenol toxic n.d.toxic n.d.

[1022] TABLE 5 Triazine compounds useful in treating inflammatorydisorders generally, and all diseases related to inhibition of IL6. %IL6 production % IL6 production Inhibition of TNF- Inhibition compare toTNF compare to AGE induced IL6 of TNF- stimulation of IL6 (100%).stimulation of IL6 (100%). Patent Name production induced IL6 Compdstested @ 10 uM Compds tested @ 10 uM Number STRUCTURE Generated byAutonom (TM) (See NOTE) production HAEC, IL6 HAEC, IL6 76

N2-cycloheptyl-N4-ethyl-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d. 18 n.d. 77

N2-(tert-butyl)-N4-cycloheptyl-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d. 21 n.d. 78

N2-benzyl-N4-cycloheptyl-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine +++ n.d. 47 n.d. 82

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-(1-pyrrolidinyl)-1,3,5-triazine-2,4-diamine +++ n.d. 34 n.d. 83

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-hexahydro-1H-azepin-1-yl-1,3,5-triazine-2,4-diamine ++++ n.d. 18 n.d. 84

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-octahydro-1(2H)-quinolinyl-1,3,5-triazine-2,4-diamine +++ n.d. 33 n.d. 85

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-(4-methylcyclohexyl)-1,3,5-triazine-2,4,6-triamine +++ n.d. n.d. 86

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-[(2S)-2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5-triazine-2,4-diamine ++++ n.d. 19 n.d. 87

N2-cycloheptyl-N4-(3-fluoro-4-methoxyphenyl)-6-(4-methyl-1-piperazinyl)- 1,3,5-triazine-2,4-diaminen.d. ++++ n.d.  5 88

6-(4-acetyl-1-piperazinyl)-N2-cycloheptyl-N4-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4-diamine ++++ n.d. 19n.d. 91

N2-cycloheptyl-N4-(3-fluoro-4-methoxyphenyl)-N6-(2-furanylmethyl)-1,3,5- triazine-2,4,6-triamine +++n.d. 28 n.d. 93

N2-cycloheptyl-N4-[2-(dimethylamino)ethyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5- triazine-2,4,6-triamine ++++ ++++13 6 (IC50 = 3.36) 94

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-{4-[2-oxo-(1-pyrrolidinyl)ethyl]-1-piperazinyl}1,3,5- triazine-2,4-diamine +++ ++++32 10 (IC50 = 5.8) 95

N-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-N″-(3-fluoro-4-methoxy-phenyl)- [1,3,5]triazine-2,4,6-triamine++++ ++++ 5 (IC50 = 2.8) 1 (IC50 = 6.6) 96

N2-cycloheptyl-N4-(3-fluoro-4-methoxyphenyl)-N6-[2-(1-piperidinyl)ethyl]-1,3,5-triazine-2,4,6-triamine ++++ n.d.  6 n.d. 99

1-[3-({4-(cycloheptylamino)-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2- yl}amino)propyl]-2-pyrrolidinone+++ ++++ 39 8 (IC = 6.04) 100

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-N6-[3-(1H-imidazol-1-yl)propyl]-1,3,5-triazine-2,4,6-triamine n.d. ++++ n.d.  1 3

N2-cycloheptyl-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(3-quinolinyl)-1,3,5- triazine-2,4,6-triaminen.d. ++++ n.d.  1 4

N2-cycloheptyl-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(6-quinolinyl)-1,3,5- triazine-2,4,6-triaminen.d. ++++ n.d.  1 10

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(4-fluorophenyl)-1,3,5-triazine-2,4,6-triamine nd. ++++ n.d.  1 11

4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]benzonitrile n.d.++++ n.d. 1 (IC50 = 8.87) 24

N2-cycloheptyl-N4-(2,3-dihydro-1,4- benzodioxin-6-yl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6- triamine n.d. ++++ n.d.  1 25

N2-cycloheptyl-N4-[4- (dimethylamino)phenyl]-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine-2,4,6- triamine n.d. ++++ n.d.  1 26

N²-(3-chloro-4-diethylamino-phenyl)-N⁴-cycloheptyl-N⁶-(1-ethyl-pyrrolidin-2-ylmethyl)-1,3,5-triazine-2,4,6-triamine ++++ n.d. 1 (over 50% of cellswere floating) n.d. 27

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(4-morpholinyl)phenyl]-1,3,5-triazine-2,4,6- triamine ++++ ++++  3  1 28

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-[4-(4-methyl-1-piperazinyl)phenyl]-1,3,5-triazine-2,4,6- triamine ++++ n.d. 23 n.d. 29

N-{4-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]phenyl}acetamide++++ ++++ 13  7 30

N-{3-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)- amino]phenyl}acetamide++++ ++++  9 17 32

N2-cycloheptyl-N4-(4-ethoxyphenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine- 2,4,6-triamine n.d. ++++n.d.  1 34

N2-cycloheptyl-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(2-pyridinyl)-1,3,5- triazine-2,4,6-triamine++++ ++++  3  1 35

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-methylphenyl)1,3,5-triazine-2,4,6-triamine +++ ++++ 34  4 39

2-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)-amino]-4-methyl-3-thiophenecarboxamide nd. +++ n.d. 28 40

N2-(4-chlorophenyl)-N4-cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N2-methyl-1,3,5- triazine-2,4,6-triaminen.d. ++++ n.d. 19 41

3-[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl(phenyl)amino]propanenitrile n.d. ++++ n.d. 18 42

N2-cycloheptyl-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(4-methoxyphenyl)-N6-methyl-1,3,5-triazine-2,4,6-triamine ++++ ++++  1  1 43

N2-cycloheptyl-N4-(2,4-difluorophenyl)-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-methyl-1,3,5-triazine-2,4,6-triamine n.d. ++++ n.d.  4 44

[(4-(cycloheptylamino)-6-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-1,3,5-triazin-2-yl)(phenyl)amino]acetonitrile n.d. ++++ n.d. 1 (IC50 = 7.1) 45

N2-(3-chlorophenyl)-N4-cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-N2-methyl-1,3,5- triazine-2,4,6-triaminen.d. ++++ n.d.  1 46

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-methyl-N6-[2-(trifluoromethyl)phenyl]-1,3,5-triazine- 2,4,6-triamine n.d. ++++ n.d. 3 48

N2-(3-chloro-4-methoxyphenyl)-N4- cycloheptyl-N6-[(1-ethyl-2-pyrrolidinyl)methyl]-1,3,5-triazine- 2,4,6-triamine +++ n.d. 3074 (IC50= 4.3) n.d. 51

N2-ethyl-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1 3,5- triazine-2,4,6-triamine ++++ n.d. 1 n.d. 52

N2-(tert-butyl)-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ +++  1 35 53

N2-benzyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ +++  1 32 55

N2-cyclohexyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d.  1 n.d. 56

N2-cyclopentyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d.  1 n.d. 57

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-(1-pyrrolidinyl)- 1,3,5-triazine-2,4-diamine++++ +++  1 25 58

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-hexahydro-1H-azepin-1-yl-1,3,5-triazine-2,4-diamine ++++ +++  1 30 60

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-N6-(4- methylcyclohexyl)-1,3,5-triazine-2,4,6-triamine ++++ n.d.  1 n.d. 61

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(4- fluoro-3-methoxyphenyl)-6-[2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5- triazine-2,4-diamine ++++ +++  238 (IC50 = 6.47) 62

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5-triazine-2,4-diamine n.d. +++ n.d. 42 63

6-(4-acetyl-1-piperazinyl)-N2-[(1-ethyl-2-pyrrolidinyl)methyl-N4-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine +++ n.d. 26 n.d. 64

4-{4-{[(1-ethyl-2-pyrrolidinyl)methyl]amino}-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2-yl}-1-piperazinecarboxylate +++ n.d. 23 n.d. 65

N2-(cyclohexylmethyl)-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ +++ 1 (IC50 = 6.7) 45(IC50 = 3.24) 66

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-N6-(2-furylmethyl)-1,3,5-triazine-2,4,6-triamine ++++ ++++  1 23 67

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-N6-(2,2,2-trifluoroethyl)-1,3,5-triazine-2,4,6-triamine n.d. +++ n.d. 39 68

N2-[2-(dimethylamino)ethyl]-N4-[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ +++  1 40 (IC50 =7.7) 69

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-{4-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1-piperazinyl}-1,3,5- triazine-2,4-diamine ++++ ++++ 1  5 70

N2,N4-bis[(1-ethyl-2-pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine- 2,4,6-triamine ++++ +++  1 3571

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-N6-[2-(1-piperidinyl)ethyl]-1,3,5-triazine-2,4,6- triamine ++++ +++  1 29 72

6-[4-(1,3-benzodioxol-5-ylmethyl)-1- piperazinyl]-N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4-diamine ++++ +++  5 31 73

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-[4-(2-pyridinyl)-1-piperazinyl]-1,3,5-triazine-2,4-diamine ++++ ++++ 1 (IC50 = 8.7) 20(IC50 = 7.5) 74

1-[3-({4-{[(1-ethyl-2- pyrrolidinyl)methyl]amino}-6-[(3-fluoro-4-methoxyphenyl)amino]-1,3,5-triazin-2- yl}amino)propyl]-2-pyrrolidinonen.d. +++ n.d. 49 75

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-N6-[3-(1H-imidazol-1-yl)propyl]-1,3,5-triazine-2,4,6- triamine +++ +++ 43 46 140

N²-cycloheptyl-N⁴-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)- 1,3,5-triazine-2,4,6-triamine++++ n.d.   12.57 n.d. 139

N²-cycloheptyl-N⁴-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)- 1,3,5-triazine-2,4,6-triamine++++ n.d.   16.08 n.d. 127

6-Chloro-N-cycloheheptyl-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine +++ n.d. 48 n.d. 141

N²-cyclohexylmethyl-N⁴-((S)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d.   23.44 n.d. 142

N²-cyclohexylmethyl-N⁴-((R)-1-ethyl-pyrrolidin-2-ylmethyl)-N⁶-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++++ n.d.   21.62 n.d. 143

({4-cycloheptylamino-6-[((S)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]-1,3,5-triazin-2-yl}-phenyl-amino)-acetonitrile ++++ n.d.   18.88 n.d. 144

({4-cycloheptylamino-6-[((R)-1-ethyl-pyrrolidin-2-ylmethyl)-amino]1,3,5-triazin-2-yl)-phenyl-amino)-acetonitrile ++++ n.d. 157

N,N′-dicyclopropyl-N″-(3-fluoro-4-methoxy-phenyl)-1,3,5-triazine-2,4,6-triamine ++ n.d.   60.48 n.d. 160

N²-cycloheptyl-N⁴-(3-fluoro-4- methoxyphenyl)-N⁶-methyl-N⁶-(1-methyl-piperidin-4-yl)-1,3,5-triazine-2,4,6-triamine, hydrogen chloride salt++++ n.d.  1 n.d. 101

(3-Chloro-4-methoxy-phenyl)-(4,6-dichloro- [1,3,5]triazin-2-yl)-aminen.d. n.d. n.d. n.d. 107

N²-(3-chloro-4-methoxy-phenyl)-N⁴-isopropyl-N⁶-methyl-N⁶-piperidin-4-yl-1,3,5- triazine-2,4,6-triamine ++n.d. 60 @ 5 uM n.d. 108

5-{4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino}-pentan-1-ol +++ n.d. 25 n.d. 109

5-[4-(3-chloro-4-methoxy-phenylamino)-6-(methyl-piperidin-4-yl-amino)-1,3,5-triazin-2- ylamino]-pentan-1-ol +++n.d. 40 n.d. 111

N-Butyl-N′-(3-chloro-4-methoxy-phenyl)-N′(1-methyl-piperidin-4-yl)-N-propyl-[1,3,5]triazine-2,4,6-triamine + n.d. >90 [46 @ 5 uM] did not seetoxicity n.d. 112

N²-Butyl-N⁴-(3-chloro-4-methoxy-phenyl)-N⁶-methyl-N⁶-piperidin-4-yl-N²-propyl-1,3,5- triazine-2,4,6-triamine +n.d. 90 @ 5 uM did not see toxicity n.d. # production. “n.d.” means thatthe activity of the compound was not determined in the given assay.

[1023] TABLE 6 Inhibition of TNF-induced heparanase secretion PatentName Relative # STRUCTURE Generated by Autonom ™ Activity Activity 76

N2-cycloheptyl-N4-ethyl-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine ++ 50% inhibition @ 5 uM82

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-(1-pyrrolidinyl)-1,3,5-triazine-2,4-diamine +++ 73% inhibition @ 5 uM 87

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-(4-methyl-1-piperazinyl)-1,3,5-triazine-2,4-diamine + 40-60% inhibition @ 10 uM 94

N2-cycloheptyl-N4-(3-fluoro-4- methoxyphenyl)-6-{4-[2-oxo-(1-pyrrolidinyl)ethyl]-1-piperazinyl}-1,3,5- triazine-2,4-diamine + 40-60%inhibition @ 10 uM 35

N2-cycloheptyl-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(2-methylphenyl)-1,3,5- triazine-2,4,6-triamine ++ 53% inhibition @ 5 uM 52

N2-(tert-butyl)-N4-[(1-ethyl-2- pyrrolidinyl)methyl]-N6-(3-fluoro-4-methoxyphenyl)-1,3,5-triazine-2,4,6- triamine + 40-60% inhibition @ 10uM 58

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4-(3-fluoro-4-methoxyphenyl)-6-hexahydro-1H-azepin-1-yl-1,3,5-triazine-2,4-diamine + 40-60% inhibition @ 10 uM 61

N2-[(1-ethyl-2-pyrrolidinyl)methyl]-N4- (3-fluoro-4-methoxyphenyl)-6-[2-(methoxymethyl)-1-pyrrolidinyl]-1,3,5- triazine-2,4-diamine + 40-60%inhibition @ 10 uM

[1024] TABLE 7 Triazine compounds useful in treating inflammatorydisorders generally, and all diseases related to inhibition of IL6. %IL6 production compare to % IL6 production compare to Inhibition of TNF-Inhibition of TNF- TNF stimulation of IL6 (100%). AGE stimulation of IL6(100%). Compound Name (IUPAC- induced IL6 induced IL6 Compds tested @ 10uM Compds tested @ 10 uM Number STRUCTURE Generated by Autonom)production production HAEC, IL6 HAEC, IL6 172

N-Cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl- [1,3,5]triazine-2,4,6-triamine +++++ 18 n.d. 171

N-Cycloheptyl-N′-(3-fluoro-4-methoxy- phenyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine- 2,4,6-triamine +++ ++ 21 n.d. 173

[4-(4-Benzyl-piperazin-1-yl)-6- morpholin-4-yl-[1,3,5]triazin-2-yl]-(4-methoxy-phenyl)-amine ++ 47 n.d. 174

N-Cycloheptyl-6-morpholin-4-yl-N′- naphthalen-2-yl-[1,3,5]triazine-2,4-diamine +++ 34 n.d. 175

N-Cycloheptyl-N′-(3-fluoro-4-methoxy- phenyl)-6-morpholin-4-yl-[1,3,5]triazine-2,4-diamine +++ 18 n.d. 176

N-Cycloheptyl-6-morpholin-4-yl-N′- phenyl-[1,3,5]triazine-2,4-diamine+++ 33 n.d. 177

N-Cycloheptyl-N′-(4-methoxy-phenyl)- 6-morpholin-4-yl-[1,3,5]triazine-2,4-diamine +++ ++ n.d. 178

N-Benzyl-N′-cycloheptyl-N″-(4- methoxy-phenyl)-N-methyl-[1,3,5]triazine-2,4,6-triamine ++ 19 n.d. 179

N-(2-[1,3]Dioxolan-2-yl-ethyl)-N′- methyl-N′-(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl-[1,3,5]triazine- 2,4,6-triamine ++ + 180

N-Cyclopropyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-N″-naphthalen-2-yl- [1,3,5]triazine-2,4,6-triamine +++ +n.d. 5 # IL6 production in the presence of AGE or TNF.

[1025] TABLE 8 Bioactivity Data for Representative Compounds Cmpd No.Structure Proliferation Data Inflammation Data

Did not pass 5 & 10 uM screens; rounding Not Active

Did not pass 5 & 10 uM screens; blunting and rounding total cell deathat 10 uM (75% inhibition); 25% cell death at 5 uM (not active)

Not active at 5 & 10 uM 48% inhibition at 10 uM with 20% cell death

Not active at 5 & 10 uM complete inhibition at 10 uM

Not active <25% inhibition at 10 uM

Not active at 5 & 10 uM 75% inhibition at 10 uM

4.9 uM average IC50 60% inhibition at 10 uM with 20% cell death

1.85 uM average IC50 85% inhibition at 10 uM with 20% cell death

5.25 uM average IC50 total cell death at 10 uM (complete inhibition)

rounding at 5 & 10 uM total cell death at 10 uM (95% inhibition)

Not active Not Active

Not active Not Active

Not active Not Active

Not active 60% inhibition at 10 uM

6.37 uM; cell morphology change at 5 & 10 uM; no cell death Not activeat 1 or 5 uM; 30% cell death at 5 uM and total death at 10 uM

6.1 uM; cell morphology change at 7.5 uM and cell death at 10 uM totalcell death at 10 uM; 58% inhibition at 5 uM, but may be due to celldeath(50% death), not active at 1 uM

5.5 uM total cell death at 10 uM; 50% inhibition at 5 uM with 20% celldeath

Not active 35% inhibition at 10 uM

Not active 25% inhibition at 5 uM and 10 uM

3.3 uM; 4.5 uM; cell blunting at 3 to 10 uM total cell death at 10 uM(84% inhibition); 64% inhibition at 5 uM

1.5 uM; 4.2 uM; cell blunting at 3 uM and rounding at 5 uM total celldeath at 10 uM (97% inhibition); 43% inhibition at 5 uM with 25% celldeath

1.9 uM; cell blunting at 1 uM and rounding at 5 uM 41% inhibition at 10uM with >50% cell death; 37% inhibition at 5 uM with 20% cell death

6.6 uM 88% inhibition at 10 uM with >50% cell death; 69% inhibition at 5uM with 20% cell death

Not active 36% inhibition at 10 uM

Not active 25% inhibition at 10 uM

Not active 23% inhibition at 10 uM

Not tested Not tested

4.1 uM Not active with 15% cell death at 10 uM

Not active 20% inhibition at 10 uM, no effect at 5 uM

7.8 uM; 5.3 uM 35% inhibition at 10 uM, 25% inhibition at 5 uM

We claim:
 1. A compound of the formula Id:

wherein G is selected from NH or O; Z is selected from H or,

wherein X¹ is selected from F or Cl, and X² is selected from OCH₃, NH₂,OC(O)CH₃, or OH; A is selected from NR¹ or O; Y¹ is selected from R¹;

B is selected from NR¹ or O; and Y² is selected from

wherein q is 0 or 1, E is selected from O or NR² wherein R² is selectedfrom R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂; or

wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and whereinR¹ is in each occurrence independently selected from H, or a linear orbranched alkyl with up to 10 carbon atoms.
 2. A compound as claimed inclaim 1, wherein the compound is6-(3-Chloro-4-methoxy-phenoxy)-N-cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-6-(3-fluoro-4-methoxy-phenoxy)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-6-cycloheptyloxy-N′-methyl-N′-(1-methylpiperidin-4-yl)-[1,3,5]triazine-2,4-diamine,6-Cycloheptyloxy-N-(3-fluoro-4-methoxy-phenyl)-N′-methyl-N′-(1-methylpiperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-N′(3-fluoro-4-methoxy-phenyl)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamineN-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine.N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine.(3-Chloro-4-methoxy-phenyl)-[4-cycloheptyloxy-6-(1-methyl-piperidin-4yloxy)-[1,3,5]triazin-2-yl]-amine,[4-Cycloheptyloxy-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine,N-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazine-2,4-diamine,6-(3-Chloro-4-methoxy-phenoxy)-N-cycloheptyl-N′-(1-ethyl-pyrrolidin-2ylmethyl)-[1,3,5]triazine-2,4-diamine,6-Cycloheptyloxy-N-(1-ethyl-pyrrolidin-2-ylmethyl)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-6-cycloheptyloxy-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-[1,3,5]triazine-2,4-diamine,[4-Cycloheptyloxy-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]methyl-(1-methyl-piperidin-4-yl)-diamine,[4-Cycloheptyloxy-6-(piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine,Cycloheptyl-[4-(3-fluoro-4-methoxyphenoxy)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-amine,2-Cycloheptyloxy-4-(3-fluoro-4-methoxy-phenoxy)-6-(1-methyl-piperidin4-yloxy)-[1,3,5]triazine,2-(3-Chloro-4-methoxy-phenoxy)-4-cycloheptyloxy-6-(1-methyl-piperidin4-yloxy)-[1,3,5]triazine,[4-Cycloheptyloxy-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]-(1-ethyl-pyrrolidin-2-ylmethyl)-amine,[4-Cycloheptyloxy-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-[1,3,5]triazin-2yl]-(3-fluoro-4-methoxy-phenyl)-amine,Cycloheptyl-[4-(1-ethyl-pyrrolidin-2-ylmethoxy)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]-amine,2-Cycloheptyloxy-4-(1-ethyl-pyrrolidin-2-ylmethoxy)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazine,N-(4-Amino-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,1-{4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidin-1-yl}-ethanone,4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidine-1-carboxylicacid,4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidine-1-carboxylicacid amide,4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidin-1-ol,2{[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-methyl)-pyrrolidine-1-carboxylicacid,2-{[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-methyl}-pyrrolidine-1-carboxylicacid amide, Acetic acid2-chloro-4{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino)-phenylester,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-(tetrahydro-pyran-4-yI)-[1,3,5]triazine-2,4,6-triamine,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-methyl-N″-(tetrahydro-pyran-4-yl)-[1,3,5]triazine-2,4,6-triamine,4-[4-Cycloheptylamino-6-(piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]-2-fluoro-phenol,2-Chloro-4-[4-cycloheptylamino-6-(piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]-phenol,N-(4-Aminomethyl-cyclohexyl)-N′-cycloheptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine,N-(4-Aminomethyl-cyclohexyl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine,4-[4-(4-Aminomethyl-cyclohexylamino-6-cycloheptylamino-[1,3,5]triazin-2-ylamino]-2-fluoro-phenol,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-piperidin-4-ylmethyl[1,3,5]triazine-2,4,6-triamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-piperidin-4-ylmethyl[1,3,5]triazine-2,4,6-triamine,4-{4-Cycloheptylamino-6-[(piperidin-4-ylmethyl)-amino]-[1,3,5]triazin-2-ylamino}-2-fluoro-phenol,N-Cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,N-Cycloheptyl-N′-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine,N-(3-Fluoro-4-methoxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,4-(3-Fluoro-4-methoxy-phenylamino)-6-(methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazin-2-ol,4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,4-Cycloheptylamino-6-(methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,N-(4-Amino-cyclohexylmethyl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine,N-(4-Amino-cyclohexylnethyl)-N′-cycloheptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine,4-Cycloheptylamino-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol,4-[(1-Ethyl-pyrrolidin-2-ylmethyl)-amino]-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-[(1-ethyl-pyrrolidin-2-ylmethyl)amino]-(1,3,5]triazin-2-ol,4-(Cyclohexylmethyl-amino)-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,or4-(Cyclohexylmethyl-amino)-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol.3. A composition comprising a compound of the formula Id:

wherein G is selected from NH or O; Z is selected from H or

wherein X¹ is selected from F or Cl, and X² is selected from OCH₃, NH₂,OC(O)CH₃, or OH; A is selected from NR¹ or O; Y¹ is selected from R¹,

B is selected from NR¹ or O; and Y² is selected from

wherein q is 0 or 1, E is selected from O or NR² wherein R² is selectedfrom R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂; or

wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and whereinR¹ is in each occurrence independently selected from H, or a linear orbranched alkyl with up to 10 carbon atoms.
 4. The composition as claimedin claim 3, wherein the compound is selected from:6-(3-Chloro-4-methoxy-phenoxy)-N-cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-6-(3-fluoro-4-methoxy-phenoxy)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-6-cycloheptyloxy-N′-methyl-N′-(1-methylpiperidin-4-yl)-[1,3,5]triazine-2,4-diamine,6-Cycloheptyloxy-N-(3-fluoro-4-methoxy-phenyl)-N′-methyl-N′-(1-methylpiperidin-4-yl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(piperidin-4-yloxy)-[1,3,5]triazine-2,4-diamine,(3-Chloro-4-methoxy-phenyl)-[4-cycloheptyloxy-6-(1-methyl-piperidin-4yloxy)-[1,3,5]triazin-2-yl]-amine,[4-Cycloheptyloxy-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine,N-Cycloheptyl-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazine-2,4-diamine,6-(3-Chloro-4-methoxy-phenoxy)-N-cycloheptyl-N′-(1-ethyl-pyrrolidin-2ylmethyl)-[1,3,5]triazine-2,4-diamine,6-Cycloheptyloxy-N-(1-ethyl-pyrrolidin-2-ylmethyl)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-6-cycloheptyloxy-N′-(1-ethyl-pyrrolidin-2-ylmethyl)-[1,3,5]triazine-2,4-diamine,N-Cycloheptyl-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-N′-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4-diamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-[1,3,5]triazine-2,4-diamine,[4-Cycloheptyloxy-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]methyl-(1-methyl-piperidin-4-yl)-diamine,[4-Cycloheptyloxy-6-(piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-(3-fluoro-4-methoxy-phenyl)-amine,Cycloheptyl-[4-(3-fluoro-4-methoxyphenoxy)-6-(1-methyl-piperidin-4-yloxy)-[1,3,5]triazin-2-yl]-amine,2-Cycloheptyloxy-4-(3-fluoro-4-methoxy-phenoxy)-6-(1-methyl-piperidin4-yloxy)-[1,3,5]triazine,2-(3-Chloro-4-methoxy-phenoxy)-4-cycloheptyloxy-6-(1-methyl-piperidin4-yloxy)-[1,3,5]triazine,[4-Cycloheptyloxy-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]-(1-ethyl-pyrrolidin-2-ylmethyl)-amine,[4-Cycloheptyloxy-6-(1-ethyl-pyrrolidin-2-ylmethoxy)-[1,3,5]triazin-2yl]-(3-fluoro-4-methoxy-phenyl)-amine,Cycloheptyl-[4-(1-ethyl-pyrrolidin-2-ylmethoxy)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazin-2-yl]-amine,2-Cycloheptyloxy-4-(1-ethyl-pyrrolidin-2-ylmethoxy)-6-(3-fluoro-4-methoxy-phenoxy)-[1,3,5]triazine,N-(4-Amino-3-chloro-phenyl)-N′-cycloheptyl-N″-methyl-N″-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,1-{4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidin-1-yl}-ethanone,4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidine-1-carboxylicacid,4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidine-1-carboxylicacid amide, 4-[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-piperidin-1-ol,2{[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-methyl)-pyrrolidine-1-carboxylicacid,2-{[4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ylamino]-methyl}-pyrrolidine-1-carboxylicacid amide, Acetic acid2-chloro-4{4-cycloheptylamino-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ylamino)-phenylester,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-(tetrahydro-pyran-4-yI)-[1,3,5]triazine-2,4,6-triamine,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-methyl-N″-(tetrahydro-pyran-4-yl)-[1,3,5]triazine-2,4,6-triamine,4-[4-Cycloheptylamino-6-(piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]-2-fluoro-phenol,2-Chloro-4-[4-cycloheptylamino-6-(piperidin-4-ylamino)-[1,3,5]triazin-2-ylamino]-phenol,N-(4-Aminomethyl-cyclohexyl)-N′-cycloheptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine,N-(4-Aminomethyl-cyclohexyl) -N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine,4-[4-(4-Aminomethyl-cyclohexylamino-6-cycloheptylamino-[1,3,5]triazin-2-ylamino]-2-fluoro-phenol,N-Cycloheptyl-N′-(3-fluoro-4-methoxy-phenyl)-N″-piperidin-4-ylmethyl[1,3,5]triazine-2,4,6-triamine,N-(3-Chloro-4-methoxy-phenyl)-N′-cycloheptyl-N″-piperidin-4-ylmethyl[1,3,5]triazine-2,4,6-triamine,4-{4-Cycloheptylamino-6-[(piperidin-4-ylmethyl)-amino]-[1,3,5]triazin-2-ylamino}-2-fluoro-phenol,N-Cycloheptyl-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,N-Cycloheptyl-N′-piperidin-4-yl-[1,3,5]triazine-2,4,6-triamine,N-(3-Fluoro-4-methoxy-phenyl)-N′-methyl-N′-(1-methyl-piperidin-4-yl)-[1,3,5]triazine-2,4,6-triamine,4-(3-Fluoro-4-methoxy-phenylamino)-6-(methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-[methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-cycloheptylamino-[1,3,5]triazin-2-ol,4-Cycloheptylamino-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,4-Cycloheptylamino-6-(methyl-(1-methyl-piperidin-4-yl)-amino]-[1,3,5]triazin-2-ol,N-(4-Amino-cyclohexylmethyl)-N′-(3-chloro-4-methoxy-phenyl)-N″-cycloheptyl-[1,3,5]triazine-2,4,6-triamine,N-(4-Amino-cyclohexyhethyl)-N′-cycloheptyl-N″-(3-fluoro-4-methoxy-phenyl)-[1,3,5]triazine-2,4,6-triamine,4-Cycloheptylamino-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol,4-[(1-Ethyl-pyrrolidin-2-ylmethyl)-amino]-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,4-(3-Chloro-4-methoxy-phenylamino)-6-[(1-ethyl-pyrrolidin-2-ylmethyl)amino]-(1,3,5]triazin-2-ol,4-(Cyclohexylmethyl-amino)-6-(3-fluoro-4-methoxy-phenylamino)-[1,3,5]triazin-2-ol,or4-(Cyclohexylmethyl-amino)-6-[(1-ethyl-pyrrolidin-2-ylmethyl)-amino]-[1,3,5]triazin-2-ol.5. The composition as claimed in claim 3, further comprising: apharmaceutically acceptable carrier; optionally, a pharmaceuticallyacceptable auxiliary; optionally, a pharmaceutically acceptablepreservative; and optionally, a pharmaceutically acceptable excipient.6. The composition as claimed in claim 3, further comprising an agentselected from a chemotherapeutic agent, an immunosuppressive agent, acytokine, a cytotoxic agent, a nucleolytic compound, a radioactiveisotope, a receptor, a pro-drug activating enzyme, an anti-inflammatoryagent, an antirheumatic agent, a cardiovascular agent, a toxin, or anycombination thereof.
 7. The composition as claimed in claim 3, whereinthe composition is in the form of a tablet, a capsule, a cachet, apowder, a granule, a solution, a suspension, an emulsion, a bolus, alozenge, a suppository, a pessary, a tamport, a cream, a gel, a paste, afoam, a spray, an aerosol, a microcapsule, a liposome, a transdermalpatch, a pastille, a paste, or a mouthwash.
 8. A method of treatingunwanted cellular proliferation, an inflammation mediated disease, or ahyperproliferative disease, or modulating a glycosidase enzyme in ahuman or an animal comprising administering to the human or animal atherapeutically effective amount of a composition comprising a compoundof the formula Id:

wherin G is selected from NH or O; Z is selected from H or

wherein X¹ is selected from F or Cl, and X² is selected from OCH₃, NH₂,OC(O)CH₃, or OH; A is selected from NR¹ or O; Y¹ is selected from R¹,

B is selected from NR¹ or O; and Y² is selected from

wherein q is 0 or 1, E is selected from O or NR² wherein R² is selectedfrom R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂; or

wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and whereinR¹ is in each occurrence independently selected from H, or a linear orbranched alkyl with up to 10 carbon atoms.
 9. A medical devicecomprising: a drug delivering or eluting member; and a composition inaccordance with claim 3 disposed on or within the drug delivering oreluting member.
 10. The medical device as claimed in claim 9, whereinthe drug delivering or eluting member is a stent.
 11. The medical deviceas claimed in claim 9, wherein the drug delivering or eluting member isselected from a shunt, a colostomy bag attachment device, an eardrainage tube, a lead for a pace maker, a lead for an implantabledefibrillator, a suture, a staple, an anastomosis device, a vertebraldisk, a bone pin, a suture anchor, a hemostatic barrier, a clamp, ascrew, a plate, a clip, a vascular implant, a tissue adhesive, a tissuesealant, a tissue scaffold, a bone substitute, an intraluminal device, astent, or a vascular support.
 12. A microarray comprising: a geneexpression profile generated from a cell type treated with a compound ofthe formula Id:

wherein G is selected from NH or O; Z is selected from H or

wherein X¹ is selected from F or Cl, and X² is selected from OCH₃,NH_(2, OC(O)CH) ₃, or OH; A is selected from NR¹ or O; Y¹ is selectedfrom R¹,

B is selected from NR¹ or O; and Y² is selected from

wherein q is 0 or 1, E is selected from O or NR² wherein R² is selectedfrom R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂; or

wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and whereinR¹ is in each occurrence independently selected from H, or a linear orbranched alkyl with up to 10 carbon atoms.
 13. The microarray as claimedin claim 12, wherein the cell type is selected from the group of cellscomprising coronary artery endothelium, umbilical artery endothelium,umbilical vein endothelium, aortic endothelium, dermal microvascularendothelium, pulmonary artery endothelium, myometrium microvascularendothelium, keratinocyte epithelium, bronchial epithelium, mammaryepithelium, prostate epithelium, renal cortical epithelium, renalproximal tubule epithelium, small airway epithelium, renal epithelium,umbilical artery smooth muscle, neonatal dermal fibroblast, pulmonaryartery smooth muscle, dermal fibroblast, neural progenitor cells,skeletal muscle, astrocytes, aortic smooth muscle, mesangial cells,coronary artery smooth muscle, bronchial smooth muscle, uterine smoothmuscle, lung fibroblast, osteoblasts, or prostate stromal cells.
 14. Anexpression profile database comprising: a patient identifying reference;and an expression profile for the patient generated by administering tothe patient a compound of the formula Id:

wherein G is selected from NH ot O; Z is selected from H or

wherein X¹ is selected from F or Cl, and X² is selected, from OCH₃, NH₂,OC(O)CH₃, or OH; A is selected from NR¹ or O; Y¹ is selected from R¹,

B is selected from NR¹or O; and Y² is selected from

wherein q is 0 or 1, E is selected from O or NR² wherein R² is selectedfrom R¹, OR¹, C(O)R¹, C(O)OR¹, C(O)NH₂, or CH₂NH₂; or

wherein R³ is selected from R¹, C(O)R¹, C(O)OR¹, or C(O)NH₂; and whereinR¹ is in each occurrence independently selected from H, or a linear orbranched alkyl with up to 10 carbon atoms.
 15. The method as claimed inclaim 8, wherein the compound is present in the composition in an amounteffective upon administration in a daily dose, a daily sub-dose, or anyappropriate fraction thereof to the human or animal to reduce theeffects of the disease or modulate the glycosidase enzyme.
 16. Themethod as claimed in claim 8, wherein the compound is present in thecomposition in an amount effective upon administration in a daily dose,a daily sub-dose, or any appropriate fraction thereof to the human oranimal to reduce the effects of the unwanted cellular proliferation. 17.The method as claimed in claim 8, wherein the compound is present in thecomposition in an amount effective upon administration in a daily dose,a daily sub-dose, or any appropriate fraction thereof to the human oranimal to modulate the glycosidase enzyme.
 18. The method as claimed inclaim 8, wherein the compound is present in the composition in an amounteffective upon administration in a daily dose, a daily sub-dose, or anyappropriate fraction thereof to the human or animal to reduce theeffects of the inflammation mediated disease.
 19. The method as claimedin claim 8, wherein the compound is present in the composition in anamount effective upon administration in a daily dose, a daily sub-dose,or any appropriate fraction thereof to the human or animal to reduce theeffects of the hyperproliferative disease.